Heterocyclic sulfonamide inhibitors of beta amyloid production

ABSTRACT

Compounds of Formula (I),  
                 
 
wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , T, W, X, Y and Z are as defined herein are provided, together with pharmaceutically acceptable salt, hydrates and/or prodrugs thereof. Methods of using these compounds for inhibiting beta amyloid production and for treatment of Alzheimer&#39;s Disease and Down&#39;s syndrome are described

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.10/455,674, filed Jun. 5, 2003, which is a continuation of U.S. patentapplication Ser. No. 10/014,304, filed Dec. 11, 2001, now U.S. Pat. No.6,610,734, issued Aug. 26, 2003, which claims the benefit of thepriority of U.S. Provisional Patent Application No. 60/255,105, filedDec. 13, 2000, now abandoned.

BACKGROUND OF THE INVENTION

This invention relates to inhibitors of beta amyloid production, whichhave utility in the treatment of Alzheimer's disease.

Alzheimer's Disease (AD) is the most common form of dementia (loss ofmemory) in the elderly. The main pathological lesions of AD found in thebrain consist of extracellular deposits of beta amyloid protein in theform of plaques and angiopathy and intracellular neurofibrillary tanglesof aggregated hyperphosphorylated tau protein. Recent evidence hasrevealed that elevated beta amyloid levels in brain not only precede taupathology but also correlate with cognitive decline. Further suggestinga causative role for beta amyloid in AD, recent studies have shown thataggregated beta amyloid is toxic to neurons in cell culture.

Beta amyloid protein is composed mainly of 39-42 amino acid peptides andis produced from a larger precursor protein called amyloid precursorprotein (APP) by the sequential action of the proteases beta and gammasecretase. Although rare, cases of early onset AD have been attributedto genetic mutations in APP that lead to an overproduction of eithertotal beta amyloid protein or its more aggregation-prone 42 amino acidisoform. Furthermore, people with Down's Syndrome possess an extrachromosome that contains the gene that encodes APP and thus haveelevated beta amyloid levels and invariably develop AD later in life.

There continues to be an unmet need for compositions useful ininhibiting beta amyloid production and in the treatment of the effectsof Alzheimer's Disease (AD).

SUMMARY OF THE INVENTION

The present invention provides heterocyclic sulfonamide derivatives of2- amino-1-alcohols and related homologs that have been found tospecifically inhibit the production of beta amyloid protein from APP andto be capable of passing through the blood-brain barrier. Thesecompounds are useful for the treatment of conditions in which betaamyloid levels are elevated (e.g., AD, Down's Syndrome). Systemicadministration of these compounds to subjects at risk of, or sufferingfrom, these diseases lowers beta amyloid protein levels with subsequentreduction in the toxic beta amyloid aggregates in the brains of thesepatients.

In one aspect, the present invention provides a compound of Formula (I),as defined herein, pharmaceutically acceptable salts, hydrates, orprodrugs thereof. In one embodiment, the compounds of Formula (I) arethiophenesulfonamides. In another embodiment, the compounds of Formula(I) are furansulfonamides. Among the particularly desirable compoundsare those having a halogen in the 5-position of the heterocycle (e.g.,5-halo thiophenesulfonamides) and β-branches in the side chain of theprimary alcohol.

In another aspect, the invention provides a pharmaceutical compositioncontaining one or more compounds of Formula (I) and a physiologicallycompatible carrier.

In yet another aspect, the invention provides a method of inhibitingbeta amyloid production in-a subject by delivering a compound of Formula(I).

In still another aspect, the invention provides a method of treatingAlzheimer's Disease (AD) in a subject by administering a compound ofFormula (I) to the subject in an amount sufficient to alleviate thesymptoms or progress of AD.

These and other aspects of the invention will be apparent to one ofskill in the art upon reading of the following detailed description ofthe invention.

DETAILED DESCRIPTION OF THE INVENTION

The invention consists of compounds of Formula (I), their pharmaceuticalformulations, and their use in modulating beta amyloid production insubjects at risk for, or suffering from, AD or other diseases resultingfrom elevated levels of beta amyloid protein in the brain. The compoundsof Formula (I) include pharmaceutically acceptable salts and/or hydratesor prodrugs thereof, wherein:

-   -   R₁ and R₂ are independently selected from the group consisting        of hydrogen, alkyl, substituted alkyl, CF₃, alkenyl, substituted        alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted        cycloalkyl, phenyl, substituted phenyl, and        (CH₂)_(n)(1,3)dioxane, where n is 2 to 5;    -   R₃ is selected from the group consisting of hydrogen, alkyl, and        substituted alkyl;    -   R₄ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, alkylcycloalkyl, substituted alkylcycloalkyl,        phenyl(substituted)alkyl, alkylOH, substituted alkylOH,        alkylOBn, substituted alkylOBn, alkylpyridyl, substituted        alkylpyridyl, alkylfuranyl, substituted alkylfuranyl,        CH(OH)phenyl, CH(OH)substituted phenyl, alkenyl, substituted        alkenyl, cycloalkyl, substituted cycloalkyl,        N-substituted-piperidinyl, piperidinyl, substituted piperidinyl,        tetrahydrothiopyran, substituted tetrahydrothiopyran, 2-indane,        substituted 2-indane, phenyl, substituted phenyl, alkylNHR₇, and        substituted alkylNHR₇;    -   with the proviso that R₃ and R₄ are not both hydrogen;    -   R₇ is alkyl, substituted alkyl, cycloalkyl, substituted        cycloalkyl, benzyl, substituted benzyl, alkylOH, substituted        alkylOH, alkylSR₈, or substituted alkylSR₈;    -   R₈ is alkyl, substituted alkyl, benzyl, or substituted benzyl;    -   or R₃ and R₄ may be joined to form a ring;    -   R₅ is selected from the group consisting of hydrogen, lower        alkyl, substituted lower alkyl, alkenyl, substituted alkenyl,        alkynyl, substituted alkynyl, CH₂cycloalkyl, substituted        CH₂cycloalkyl, benzyl, substituted benzyl, and CH₂CH₂QR₉;    -   Q is O, NH or S;    -   R₉ is lower alkyl, substituted lower alkyl, phenyl, or        substituted phenyl;    -   R₆ is selected from the group consisting of hydrogen, halogen        and CF₃;    -   T is selected from the group consisting of    -   W, Y and Z are independently selected from the group consisting        of C, CR₁₀ and N;    -   R₁₀ is selected from the group consisting of hydrogen and        halogen, with the proviso that at least one of W, Y and Z must        be C;    -   X is selected from the group consisting of O, S, SO₂, and NR₁₁;    -   R₁₁ is selected from the group consisting of hydrogen, lower        alkyl, substituted lower alkyl, benzyl, substituted benzyl,        phenyl, and substituted phenyl;    -   provided that when the compound contains one or more chiral        centers, at least one of the chiral centers must be of        S-stereochemistry.

The point of attachment of the W—X—Y-Z-C heterocyclic ring to the SO₂group is not a limitation of the present invention. However, in onepreferred embodiment, the ring is attached to the SO₂ group through acarbon-atom. However, the ring may be attached through O, S, or Nheteroatoms.

The compounds of the invention may contain one or more asymmetric carbonatoms and some of the compounds may contain one or more asymmetric(chiral) centers and may thus give rise to optical isomers anddiastereomers. While shown without respect to stereochemistry in Formula(I), when the compounds of Formula (I) contain one or more chiralcenters, at least one of the chiral centers is of S-stereochemistry.Most preferably, the carbon atom to which N, T, R₃ and R₄ are attachedis of S-stereochemistry. Thus, the invention includes such opticalisomers and disastereomers; as well as the racemic and resolved,enantiomerically pure stereoisomers; as well as other mixtures of the Rand S stereoisomers, and pharmaceutically acceptable salts, hydrates,and prodrugs thereof.

The term “alkyl” is used herein to refer to both straight- andbranched-chain saturated aliphatic hydrocarbon groups having one to tencarbon atoms, preferably one to eight carbon atoms and, most preferably,one to six carbon atoms; as used herein, the term “lower alkyl” refersto straight- and branched-chain saturated aliphatic hydrocarbon groupshaving one to six carbon atoms; “alkenyl” is intended to include bothstraight- and branched-chain alkyl group with at least one carbon-carbondouble bond and two to eight carbon atoms, preferably two to six carbonatoms; “alkynyl” group is intended to cover both straight- andbranched-chain alkyl groups with at least one carbon-carbon triple bondand two to eight carbon atoms, preferably two to six carbon atoms.

The terms “substituted alkyl”, “substituted alkenyl”, and “substitutedalkynyl” refer to alkyl, alkenyl, and alkynyl as just described havingfrom one to three substituents selected from the group includinghalogen, CN, OH, NO₂, amino, aryl, heterocyclic, substituted aryl,substituted heterocyclic, alkoxy, substituted alkoxy, aryloxy,substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, arylthio.These substituents may be attached to any carbon of an alkyl, alkenyl,or alkynyl group provided that the attachment constitutes a stablechemical moiety.

The term “aryl” is used herein to refer to a carbocyclic aromaticsystem, which may be a single ring, or multiple aromatic rings fused orlinked together as such that at least one part of the fused or linkedrings forms the conjugated aromatic system. The aryl groups include, butare not limited to, phenyl, naphthyl, biphenyl, anthryl,tetrahydronaphthyl, phenanthryl, and indane.

The term “substituted aryl” refers to aryl as just defined having one tofour substituents from the group including halogen, CN, OH, NO₂, amino,alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, substitutedalkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio.

The term “substituted benzyl” refers to a benzyl group, havingsubstituted on the benzene ring, one to five substituents from the groupincluding halogen, CN, OH, NO₂, amino, alkyl, cycloalkyl, alkenyl,alkynyl, alkoxy, aryloxy, substituted alkyloxy, alkylcarbonyl,alkylcarboxy, alkylamino, and arylthio.

The term “heterocyclic” is used herein to describe a stable 4- to7-membered monocyclic or a stable multicyclic heterocyclic ring which issaturated, partially unsaturated, or unsaturated, and which consists ofcarbon atoms and from one to four heteroatoms selected from the groupincluding N, O, and S atoms. The N and S atoms may be oxidized. Theheterocyclic ring also includes any multicyclic ring in which any ofabove defined heterocyclic rings is fused to an aryl ring. Theheterocyclic ring may be attached at any heteroatom or carbon atomprovided the resultant structure is chemically stable. Such heterocyclicgroups include, for example, tetrahydrofuran, piperidinyl, piperazinyl,2-oxopiperidinyl, azepinyl, pyrrolidinyl, imidazolyl, pyridyl,pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, isoxazolyl, morpholinyl,indolyl, quinolinyl, thienyl, furyl, benzofuranyl, benzothienyl,thiamorpholinyl, thiamorpholinyl sulfoxide, isoquinolinyl, andtetrahydrothiopyran.

The term “substituted heterocyclic” is used herein to describe theheterocyclic just defined having one to four substituents selected fromthe group which includes halogen, CN, OH, NO₂, amino, alkyl, substitutedalkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, aryloxy,substituted aryloxy, alkyloxy, substituted alkyloxy, alkylcarbonyl,substituted alkylcarbonyl, alkylcarboxy, substituted alkylcarboxy,alkylamino, substituted alkylamino, arylthio, or substituted arylthio.

The term “substituted cycloalkyl” is used herein to describe acarbon-based ring having more than 3 carbon-atoms which forms a stablering and having from one to five substituents selected from the groupconsisting of halogen, CN, OH, NO₂, amino, alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, alkoxy, aryloxy, substitutedalkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, substitutedalkylamino, arylthio, heterocyclic, substituted heterocyclic,aminoalkyl, and substituted aminoalkyl.

Where the terms “substituted alkylcycloalkyl”, “substituted alkylOBn”,“substituted alkylpyridyl”, “substituted alkylfuranyl”, “substitutedalkyl NHR₇”, “substituted alkylOH”, and “substituted alkylSR₈” arerecited, the substitution may occur at the alkyl group or on thecorresponding base compound.

As used in the definition of the R₄ group, an N-substituted piperidinylgroup may be defined as are the substituted heterocyclic groups. Amongparticularly desirable substituents are N-alkyl-, N-aryl-, N-acyl-, andN-sulfonyl piperidinyl groups. One particularly suitableN-acyl-piperidinyl group is N-t-butyloxycarbonyl (BOC)-piperidine.However, other suitable substituents can be readily identified by one ofskill in the art.

The term “alkoxy” is used herein to refer to the OR group, where R isalkyl or substituted alkyl. The term “aryloxy” is used herein to referto the OR group, where R is aryl or substituted aryl. The term“alkylcarbonyl” is used herein to refer to the RCO group, where R isalkyl or substituted alkyl. The term “alkylcarboxy” is used herein torefer to the COOR group, where R is alkyl or substituted alkyl. The term“aminoalkyl” refers to both secondary and tertiary amines wherein thealkyl or substituted alkyl groups, containing one to eight carbon atoms,which may be either same or different and the point of attachment is onthe nitrogen atom.

The term “halogen” refers to Cl, Br, F, or I.

The term “ring” structure, e.g., when R₃ and R₄ may form a ringstructure, includes a monocyclic structure, a bridged cyclo structure,and fused cyclo structures, unless the type of ring structure isotherwise specified.

The compounds of the present invention can be used in the form of saltsderived from pharmaceutically or physiologically acceptable acids orbases. These salts include, but are not limited to, the following saltswith organic and inorganic acids such as acetic, lactic, citric,tartaric, succinic, fumaric, maleic, malonic, mandelic, mallic,hydrochloric, hydrobromic, phosphoric, nitric, sulfuric,methanesulfonic, toluenesulfonic and similarly known acceptable acids,and mixtures thereof. Other salts include salts with alkali metals oralkaline earth metals, such as sodium (e.g., sodium hydroxide),potassium (e.g., potassium hydroxide), calcium or magnesium.

These salts, as well as other compounds of the invention may be in theform of esters, carbamates and other conventional “pro-drug” forms,which, when administered in such form, convert to the active moiety invivo. In a currently preferred embodiment, the prodrugs are esters. See,e.g., B. Testa and J. Caldwell, “Prodrugs Revisited: The “Ad Hoc”Approach as a Complement to Ligand Design”, Medicinal Research Reviews,16(3):233-241, ed., John Wiley & Sons (1996).

In one particularly desirable-embodiment, the -compounds of Formula (I)are thiophenesulfonamides, and more desirably, 5-halothiophenesulfonamides, and most desirably, 5-halo thiophene sulfonamideswith β-branches in the side chain of a primary alcohol. Thus, withrespect to Formula (I), the compound of the invention desirably has astructure in which X is S, W is C (or CR₁₀), Y is C (or CR₁₀) and Z is C(or CR₁₀), and the sulfonamide is attached to C2 of the thiophene ring.More desirably, X is S, W is C (or CR₁₀), Y is C (or CR₁₀), Z is C (orCR₁₀) and R₆ is a halogen. Most desirably, X is S, X is C, W is C, Y isC, Z is C, R₆ is a halogen, and T is C(OH)R₁R₂, where R₁ and R₂ arehydrogen, R₃ is H, R₄ is a lower alkyl of S-stereochemistry, and R₅ isH. In preliminary screening assays in vitro and in vivo, compounds ofthese structures have been found to have unexpectedly good beta-amyloidinhibitory activity, and in many cases, better activity than compoundsof Formula (I) having other heterocycles (e.g., furans, where X is O).However, other such compounds of Formula (I) are also useful for thepurposes described herein.

For example, in another embodiment, the compounds of Formula (I) arefuransulfonamides, in which X is O, W is C, Y is C, and Z is C. In oneparticularly desirable embodiment, the furansulfonamides of Formula (I)are further characterized by β-branches in the side chain of a primaryalcohol. Thus, with respect to Formula (I), in these compounds T isC(OH)R₁R₂, in which R₁ and R₂ are hydrogen, R₃ is H, R₄ is a lower alkylof S-stereochemistry, R₅ is H and R₆ is halogen.

In still another embodiment, the compounds of Formula (I) arecharacterized by being sulfonamides of Formula (I), which haveβ-branches in the side chain of the primary alcohol group. Thus, withrespect to Formula (I), in these compounds T is C(OH)R₁R₂, R₁ and R₂ arehydrogen, R₃ is H, R₄ is a lower alkyl of S-stereochemistry, and R₅ isH.

These and the other compounds of the invention can be prepared followingthe Schemes illustrated below.

Synthesis

The compounds of the present invention can be prepared in a number ofways well known to one skilled in the art of organic synthesis. Thecompounds of the present invention can be prepared using the methodsdescribed below, together with synthetic methods known in the syntheticorganic arts or variations of these methods by one skilled in the art.(See, generally, Comprehensive Organic Synthesis, “Selectivity, Strategy& Efficiency in Modern Organic Chemistry”, ed., I. Fleming, PergamonPress, New York (1991); Comprehensive Organic Chemistry, “The Synthesisand Reactions of Organic Compounds”, ed. J. F. Stoddard, Pergamon Press,New York (1979)). Preferred methods include, but are not limited to,those outlined below.

A first method of preparation consists of reaction of a 2-aminoalcoholII with the appropriate sulfonyl halide in the presence of a base suchas triethylamine (TEA) and in a suitable solvent to afford compounds ofFormula III. For compounds where R₂ and R₁ are hydrogen, oxidation ofthe N-sulfonyl primary alcohol with pyridinium chlorochromate (PCC) orunder Swern conditions then affords the corresponding aldehyde IV whichcan be reacted with Grignard reagents (RMgX, where R is an organicradical and X is a halogen) to afford the secondary alcohols V as amixture of diastereomers which can be separated by high performanceliquid chromatography (HPLC) (Scheme 1).

A second method of preparation involves reaction of an α-amino acid orester IX with the appropriate sulfonyl halide in the presence of a basesuch as triethylamine and in a suitable solvent to afford compounds ofFormula X (Scheme 2). The intermediate N-sulfonyl acid X (Rx=H) can beconverted to the corresponding primary alcohol VIII (R₁═R₂═H) utilizingstandard methodology such as LiAlH₄, B₂H₆ or cyanuric chloride/NaBH₄.The intermediate N-sulfonyl ester X (Rx=alkyl, Bn) can also be reducedto the corresponding primary alcohol VIII (R₁═R₂═H) utilizing standardmethodology such as LiAlH₄. Alternatively, the intermediate N-sulfonylester X (Rx=alkyl, Bn) can be converted to the aldehyde IV with DiBAL.Finally, the intermediate N-sulfonyl ester X (Rx=alkyl, Bn) can bereacted with 2 equivalents of Grignard reagent to afford the tertiaryalcohols III with R₁═R₂. Alternatively, for tertiary alcohols III withR₁ not equal to R₂, the corresponding Weinreb amide (see Scheme 10) ofthe N-sulfonyl acid can be prepared and subsequently reacted with R₁MgXand R₂MgX. For compounds of formula X (Rx=H) that have an asymmetriccenter at the α-amino acid carbon, the pure enantiomers can be obtainedby standard resolution procedures employing recrystallization of saltsformed with various chiral bases.

In a variation of the second method to prepare the primary alcohols, anα-amino acid or ester (or N-protected derivative thereof) VI is firstconverted to the corresponding primary 2-aminoalcohol VII (using themethodology outlined in the previous paragraph), which is subsequently,after deprotection (if necessary), reacted with the appropriate sulfonylhalide (Scheme 3) to afford compounds of Formula VIII. For preparationof compounds derived from unnatural α-amino acids containing betabranching in the amino acid side chain, a method of preparation based onthe work of Hruby (Tet. Lett. 38: 5135-5138 (1997)) is outlined inScheme 4. This route entails formation of the α,β-unsaturated amide XIIof the Evans chiral auxiliary from an α,β-unsaturated acid XI, followedby conjugate addition of an organocuprate, trapping of the resultingenolate anion XIII with NBS, displacement of the bromide XIV with azideanion (provided by tetramethylguanidinium azide (TMGA)) to afford XV,followed by reduction to the 2-amino alcohol and subsequentsulfonylation to afford the target compound XVI. In Schemes 1 through 4,R₅ is H.

For the preparation of N-alkylated sulfonamides VIII (R₅=alkyl etc.),the sulfonamide ester XVII can be N-alkylated by either treatment with asuitable base such as potassium carbonate followed by the alkylatingagent R₅X or by employing Mitsunobu conditions (R₅OH/DEAD, TPP). LiBH₄reduction of the N-alkylated sulfonamide ester affords the N-alkylatedsulfonamide in the primary alcohol series VIII (Scheme 5). These primaryalcohols VIII can be converted to the secondary alcohols V or aldehydeIV series by chemistry that has been outlined above. Alternatively, theN-alkylated sulfonamide esters, or their corresponding Weinreb amides,can be treated with Grignard reagents to afford the N-alkylated tertiaryalcohols III.

When the heterocycle attached to the sulfonamide in the above alcoholsis thiophene, the corresponding sulfone derivative XIX may be obtainedby oxidation of the thiophene compound XVIII with MCPBA (Scheme 6).

An alternate preparation of sulfonamides derived from unnatural2-aminoalcohols utilizes the Bucherer modification of the Streckerα-amino acid synthesis (Scheme 7). In this route, an aldehyde XX isreacted with cyanide anion and ammonium carbonate to afford thehydantoin XXI, which is hydrolyzed to the α-amino acid XXII. Thiscompound is then reduced to XXIII and sulfonylated to afford the desiredcompounds of Formula XXIV.

For sulfonamides derived from 2-aminoalcohols containing an N or Oheteroatom in the side chain, a route has been devised starting fromD-serine (Scheme 8). In this route, D-serine XXV is first sulfonylatedto XXVI and subsequently converted to the ketone XXVII, which isreductively aminated to the target compounds of Formula XXVIII.

For sulfonamides derived from 2-aminoalcohols in the secondary alcoholseries with R₁═H and R₂═CF₃ (compound XXIX), a method of preparation hasbeen devised that is outlined in Scheme 9 starting from the aldehyde IV(prepared as in Scheme 1).

As has been mentioned in the section concerning Scheme 1, thepreparation of sulfonamides derived from 2-aminoalcohols in thesecondary alcohol series V results in the formation of a diastereomericmixture. An alternate method of preparation of these compounds thatresults in the production of a pure diastereomer is outlined in Scheme10 for compounds derived from L-isoleucine. This method, which utilizeschemistry previously employed by Roux (Tetrahedron 50: 5345-5360(1994)), consists of addition of Grignard reagents to the Weinreb amideXXX (derived from the requisite α-amino acid) followed by stereospecificreduction of the ketone XXXI to afford a single diastereomericN-protected 2-amino alcohol XXXII. Deprotection of this compoundfollowed by reaction with sulfonyl chlorides affords the purediastereomeric sulfonamide secondary alcohols of Formula XXXIII.

When the heterocycle attached to the sulfonamide in the above alcoholsis thiophene, the corresponding 5-iodo and 5-fluoro-thiophenederivatives may be obtained by conversation of the 5-bromo-thiophenederivative XXXIV (obtained as in Scheme 1) to a 5-trialkyltin-thiopheneintermediate XXXV which can be converted to either the 5-iodo-thiophene(XXXVII) by treatment with sodium iodide and chloramine T or the5-fluoro-thiophene analog (XXXVI) by treatment with SELECTFLUOR™(Aldrich Chemical Co) (Scheme 11).

Sulfonamides derived from cyclohexylglycinol substituted by alkoxy andamino groups at the 4 position of the cyclohexane ring can be preparedaccording to the methods described herein (Scheme 12). This routeentails initial hydrogenation of 4-L-hydroxyphenylglycine XXXVIII,followed by sulfonylation, reduction of the carboxylic acid withdiborane and formation of the N,O-acetonide XXXIX. The 4-hydroxyacetonide XXXIX is then O-alkylated using sodium hydride and analkylating agent such as an alkyl or benzyl bromride. This is followedby removal of the protecting group by treatment with aqueous acid toafford the 4-ether derivatives of Formula XXXX. Alternatively, the4-hydroxy acetonide XXXIX can be oxidized to the 4-ketone which can bereductively aminated and deprotected to afford the corresponding 4-aminoanalogs of Formula XXXXI.

Another method of preparing chirally pure N-sulfonyl 2-amino alcoholsderived from α-amino acids is outlined in Scheme 13. This methodinvolves construction of an Evans oxazolidone chiral auxiliary XXXXIIIfrom XXXXII, which is then converted to the corresponding enolate andelectrophilically aminated with trisyl azide to afford the keyintermediate XXXXIV (J. Am. Chem. Soc. 109: 6881-6883 (1987)). The azideintermediate XXXXIV is then hydrolyzed to the α-azido acid XXXXV andreduced to the chirally pure α-amino acid XXXXVI which can be convertedto the corresponding N-sulfonyl 2-amino alcohols by methods previouslydescribed above (e.g. Scheme 2).

Finally, chirally pure α-amino acids XXXXVI, one of the possiblesynthetic precursors of chiral N-sulfonyl 2-amino alcohols as mentionedabove, can also be prepared utilizing an asymmetric variant of theStrecker α-amino acid synthesis as outlined in Scheme 14 (J. Org. Chem.54:1055-1062 (1989)).

Oximes XXXIV can be derived from the corresponding aldehydes IV bystandard methodology as depicted in Scheme 15.

Methods of Use

Compounds of Formula (1) are inhibitors of beta amyloid production. Inpreliminary studies using protease specific assays, exemplary compoundsof Formula (I) have been shown to exhibit specific inhibition withrespect to protease activity. Thus, the compounds of the presentinvention are useful for treatment and prevention of a variety ofconditions in which modulation of beta amyloid levels provides atherapeutic benefit. Such conditions include, e.g., amyloid angiopathy,cerebral amyloid angiopathy, systemic amyloidosis, Alzheimer's Disease(AD), hereditary cerebral hemorrhage with amyloidosis of the Dutch type,inclusion body myositis, Down's syndrome, among others.

In addition, the compounds of Formula (I) may be utilized in generatingreagents useful in diagnosis of conditions associated with abnormallevels of beta amyloid. For example, the compounds of Formula (I) may beused to generate antibodies, which would be useful in a variety ofdiagnostic assays. Methods for generating monoclonal, polyclonal,recombinant, and synthetic antibodies or fragments thereof, are wellknown to those of skill in the art. (See, e.g., E. Mark and Padlin,“Humanization of Monoclonal Antibodies”, Chapter 4, The Handbook ofExperimental Pharmacology, Vol. 113, The Pharmacology of MonoclonalAntibodies, Springer-Verlag (June, 1994); Kohler and Milstein and themany known modifications thereof; PCT Patent Application No.PCT/GB85/00392; British Patent Application Publication No. GB2188638A;Amit et al., Science, 233:747-753 (1986); Queen et al., Proc. Nat'l.Acad. Sci. USA, 86:10029-10033 (1989); International Patent PublicationNo. WO90/07861; and Riechmann et al., Nature, 332:323-327 (1988); Huseet al, Science, 246:1275-1281 (1988)). Alternatively, the compounds ofFormula (I) may themselves be used in such diagnostic assays. Regardlessof the reagent selected (e.g., antibody or compound of Formula (I)),suitable diagnostic formats including, e.g., radioimmunoassays andenzyme-linked immunosorbent assays (ELISAs), are well known to those ofskill in the art and are not a limitation on this embodiment of theinvention.

The beta amyloid inhibitory activity of many of the compounds of thepresent invention has been determined using the Repressor Release Assay(RRA). See, Table 23 below. A compound is considered active in RRA if itleads to at least a 1.5 fold increase in luciferase activity at 20 μMand is non-toxic.

Additionally, cellular, cell-free and in vivo screening methods todetect inhibitors of beta amyloid production are known in the art. Suchassays may include radioimmunoassays and enzyme-linked immunosorbentassay (ELISA), among others. See, e.g., P. D. Mehta, et al., Techniquesin Diagnostic Pathology, vol. 2, eds., Bullock et al, Academic Press,Boston, pages 99-112 (1991), International Patent Publication No. WO98/22493, European Patent No. 0652009, U.S. Pat. No. 5,703,129 and U.S.Pat. No. 5,593,846. Selection of an appropriate in vitro or in vivoscreening assay is not a limitation of the present invention.

Pharmaceutical Formulation

The compounds of this invention may be administered to a subject by anydesirable route, taking into consideration the specific condition forwhich it has been selected. By subject is meant any suitable mammal,including humans, domestic animals (e.g., canines and felines), andlivestock, which have been recognized as having or at risk of having oneor more of the conditions for which modulation of beta amyloid levels isdesirable. Thus, the compounds of the invention are useful for treatmentand/or prevention of a number of human and veterinary conditions. Asused herein, “prevention” encompasses prevention of symptoms in asubject who has been identified as at risk for the condition, but hasnot yet been diagnosed with the same and/or who has not yet presentedany symptoms thereof.

These compounds may be delivered or administered by any suitable routeof delivery, e.g., oral, intravenous, subcutaneous, intramuscular,sublingual, intracranial, epidural, intratracheal, rectal, vaginal,among others. Most desirably, the compounds are delivered orally or by asuitable parenteral route. The compounds may be formulated incombination with conventional pharmaceutical carriers that arephysiologically compatible. Optionally, one or more of the compounds ofthe invention may be mixed with other active agents.

Suitable physiologically compatible carriers may be readily selected byone of skill in the art. For example, suitable solid carriers include,among others, one or more substances which may also act as lubricants,solubilizers, suspending agents, fillers, glidants, compression aids,binders or tablet-disintegrating agents or an encapsulating material. Inpowders, the carrier is a finely divided solid, which is in admixturewith the finely divided active ingredient. In tablets, the activeingredient is mixed with a carrier having the necessary compressionproperties in suitable proportions and compacted in the shape and sizedesired. The powders and tablets preferably contain up to 99% of theactive ingredient. Suitable solid carriers include, for example, starch,sugars (including, e.g., lactose and sucrose), dicalcium phosphate,cellulose (including, e.g., microcrystalline cellulose, methylcellulose, sodium caroboxymethyl cellulose), and kaolin.

Liquid carriers may be used in preparing solutions, suspensions,emulsions, syrups and elixirs. The active ingredient of this inventioncan be dissolved or suspended in a pharmaceutically acceptable liquidcarrier such as water, an organic solvent, a mixture of both orpharmaceutically acceptable oils or fat. The liquid carrier can containother suitable pharmaceutical additives such as solubilizers,emulsifiers, buffers, suspending agents, thickening agents, viscosityregulators, stabilizers or osmo-regulators. Suitable examples of liquidcarriers for oral and parenteral administration include water(particularly containing additives as above e.g. cellulose derivatives,preferably sodium carboxymethyl cellulose solution), alcohols (includingmonohydric alcohols and polyhydric alcohols, e.g., glycols) and theirderivatives, and oils (e.g., fractionated coconut oil, arachis oil, cornoil, peanut oil, and sesame oil). For parenteral administration thecarrier can also be an oily ester such as ethyl oleate and isopropylmyristate. Sterile liquid carriers are used in sterile liquid formcompositions for parenteral administration.

Optionally, additives customarily employed in the preparation ofpharmaceutical compositions may be included in the compositions of theinvention. Such components include, e.g., sweeteners or other flavoringagents, coloring agents, preservatives, and antioxidants, e.g., vitaminE, ascorbic acid, BHT and BHA.

Liquid pharmaceutical compositions that are sterile solutions orsuspensions can be utilized by, for example, intramuscular,intraperitoneal or subcutaneous injection. Sterile solutions can also beadministered intravenously. Oral administration may be either liquid orsolid composition form.

Preferably the pharmaceutical composition is in unit dosage form, e.g.as tablets or capsules. In such form, the composition is sub-divided inunit dose containing appropriate quantities of the active ingredient;the unit dosage forms can be packaged compositions, for example packetedpowders, vials, ampoules, prefilled syringes or sachets containingliquids. The unit dosage form can be, for example, a capsule or tabletitself, or it can be the appropriate number of any such compositions inpackage form.

As described herein, a therapeutically or prophylactically useful amountof a compound of the invention is that amount of a compound whichalleviates the symptoms of the disease, e.g., AD, or which prevents theonset of symptoms, or the onset of more severe symptoms. Generally, anindividual dose (i.e., per unit, e.g., tablet) of a compound of theinvention may be in the range from about 1 μg/kg to about 10 g/kg, morepreferably 10 mg/kg to about 5 g/kg, and most preferably about 1 mg/kgto about 200 mg/kg. Desirably, these amounts are provided on a dailybasis. However, the dosage to be used in the treatment or prevention ofa specific cognitive deficit or other condition may be subjectivelydetermined by the attending physician. The variables involved includethe specific cognitive deficit and the size, age and response pattern ofthe patient. For example, based upon the activity profile and potency ofthe compounds of this invention, a starting dose of about 10 mg per daywith gradual increase in the daily dose to about 200 mg per day mayprovide the desired dosage level in the human.

Alternatively, the use of sustained delivery devices may be desirable,in order to avoid the necessity for the patient to take medications on adaily basis. “Sustained delivery” is defined as delaying the release ofan active agent, i.e., a compound of the invention, until afterplacement in a delivery environment, followed by a sustained release ofthe agent at a later time. Those of skill in the art know suitablesustained-delivery devices. Examples of suitable sustained deliverydevices include, e.g., hydrogels (see, e.g., U.S. Pat. Nos. 5,266,325;4,959,217; and 5,292,515), an osmotic pump, such as described by Alza(U.S. Pat. No. 4,295,987 and U.S. Pat. No. 5,273,752) or Merck (EuropeanPatent No. 314,206), among others; hydrophobic membrane materials, suchas ethylenemethacrylate (EMA) and ethylenevinylacetate (EVA);bioresorbable polymer systems (see, e.g., International PatentPublication No. WO 98/44964, Bioxid and Cellomeda; U.S. Pat. No.5,756,127 and U.S. Pat. No. 5,854,388); other bioresorbable implantdevices have been described as being composed of, for example,polyesters, polyanhydrides, or lactic acid/glycolic acid copolymers(see, e.g., U.S. Pat. No. 5,817,343 (Alkermes Inc.)). For use in suchsustained delivery devices, the compounds of the invention may beformulated as described herein.

EXAMPLES

The following examples are provided to illustrate the production andactivity of representative compounds of the invention and to illustratetheir performance in a screening assay. One skilled in the art willappreciate that although specific reagents and conditions are outlinedin the following examples, these reagents and conditions are not alimitation on the present invention.

Example 13-Bromo-5-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]thiophene-2-sulfonamide

To a solution of (S)-+-isoleucinol (23 mg, 0.2 mmol) in THF (3 mL) wasadded triethylamine (46 μL, 0.24 mmol) and3-bromo-5-chlorothiophene-2-sulfonyl chloride (59.2 mg, 0.2 mmol). Thesolution was stirred for 8-16 h, then concentrated. The residue wasdissolved in MeOH (1.5 mL) and purified by semi-preparative RP-HPLC¹ togive Example 1 (20.3 mg).

The following compounds (Examples 1-7, Table 1) were prepared using3-bromo-5-chlorothiophene-2-sulfonyl chloride,5-bromothiophene-2-sulfonyl chloride,3-bromo-2-chlorothiophene-5-sulfonyl chloride,5-chlorothiophene-2-sulfonyl chloride, 2,5-dichlorothiophene-3-sulfonylchloride, 2,3-dichlorothiophene-5-sulfonyl chloride, and2-thiophenesulfonyl chloride and following the procedure outlined inExample 1. TABLE 1

(LCMS² Data: Molecular ion and retention time) RSO₂Cl(S)-(+)-isoleucinol 3-bromo-5-chlorothiophene-2-sulfonyl Example 1chloride (377 M + H); 3.25 min 5-bromothiophene-2-sulfonyl chlorideExample 2 (344 M + H); 3.01 min 3-bromo-2-chlorothiophene-s-sulfonylExample 3 chloride (378 M + H); 3.35 min 5-chlorothiophene-2-sulfonylchloride Example 4 (298 M + H); 2.97 min2,5-dichlorothiophene-3-sulfonyl chloride Example 5 (332 M + H); 3.18min 2,3-dichlorothiophene-5-sulfonyl chloride Example 6 (332 M + H);3.33 min 2-thiophenesulfonyl chloride Example 7 (264 M + H); 2.35 min

Example 85-Chloro-N-[(1S)-1-(hydroxymethyl)-2-methylpropyl]thiophene-2-sulfonamide

To a solution of L-valinol (25.8 mg, 0.25 mmol) in THF (3 mL) was addedtriethylamine (58 μL, 0.3 mmol) and 5-chlorothiophene-2-sulfonylchloride (54 mg, 0.25 mmol). The solution was stirred for 8 to 16 h,then concentrated. The residue was dissolved in MeOH (1.5 mL) andpurified by semi-preparative RP-HPLC¹ to give Example 8(19.5 mg).

The following compounds (Examples 8-10, Table 2) were prepared using5-thiophene-2-sulfonyl chloride and 5-bromothiophenesulfonyl chloridewith L-valinol and D-valinol and following the procedure outlined inExample 8. TABLE 2

(LCMS² Data: Molecular ion and retention time) RSO₂Cl5-chlorothiophene-2-sulfonyl 5-bromothiophene-2-sulfonyl Amine chloridechloride L-valinol Example 8 Example 9 (284 M + H); 2.70 min (330 M +H); 2.75 min D-valinol Example 10 (330 M + H); 2.75 min

Example 114,5-Dibromo-N-[(1S)-1-(hydroxymethyl)-2-methylpropyl]thiophene-2-sulfonamide

To a solution of (S)-(+)-2-amino-3-methyl-1-butanol (20.6 mg, 0.2 mmol)in THF (3 mL) was added triethylamine (46 μL, 0.24 mmol) and4,5-dibromothiophene-2-sulfonyl chloride (68 mg, 0.2 mmol). The solutionwas stirred for 8 to 16 h, the solvent was removed and residue purifiedby RP-HPLC¹ to give Example 11 (49.6 mg). TABLE 3

(LCMS² Data: Molecular ion and retention time) NH₂CH(R₁)CH₂OH RSO₂Cl(S)-(+)-2-amino-3-methyl-1-butanol 4,5-dibromothiophene-2- Example 11sulfonyl chloride (408 M + H); 3.22 min

Example 125-Chloro-N-[(1S)-1-cyclohexyl-2-hydroxyethyl]thiophene-2-sulfonamide

A. Part 1

To a solution of L-cyclohexyl-glycine (48.5 mg, 0.25 mmol) in THF (2 mL)was added lithium aluminum hydride (1 M solution in THF) (0.8 mL, 0.8mmol) and the solution heated at 60° C. for 4 h. The solution wasstirred at 25° C. for 8 to 16 hours. The reaction was quenched byaddition of water (45 μL), 15% aqueous sodium hydroxide (45 μL) andwater (105 μL) with vigorous stirring between each addition. The mixturewas then filtered and concentrated.

B. Part 2

To a solution of the residue from Part 1 in THF (3 mL) was addedtriethylamine (69 μL, 0.50 mmol) and 5-chlorothiophene-2-sulfonylchloride (54.3 mg, 0.25 mmol). The solution was stirred for 8 to 16 h,the solvent was removed and residue purified by RP-HPLC¹ to give Example12 (25.9 mg).

The following compounds (Examples 12-17, Table 4) were prepared using5-chlorothiophene-2-sulfonyl chloride, and 5-bromothiophene-2-sulfonylchloride with L-cyclohexylglycine, β-methyl-DL-phenylalanine, andL-allo-isoleucine and following the procedure outlined in Example 12.TABLE 4

(LCMS² Data: Molecular ion and retention time) RSO₂Cl5-chlorothiophene-2- 5-bromothiophene-2-sulfonyl Amino acid sulfonylchloride chloride L-cyclohexyl- Example 12 Example 13 glycine (324 M +H); 3.07 min (370 M + H); 3.10 min beta-methyl- Example 14 Example 15DL- (346 M + H); 3.05 min (392 M + H); 3.08 min phenylalanine L-allo-Example 16 Example 17 isoleucine (298 M + H); 2.78 min (344 M + H); 2.82min

Example 185-Bromo-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]thiophene-2-sulfonamide1,1-dioxide

A. Part 1.

To a solution of (S)-+-isoleucinol (58.6 mg, 0.5 mmol) in DCM (5 mL) wasadded triethylamine (210 μL, 1.5 mmol) and 5-bromothiophene-2-sulfonylchloride (130.8 mg, 0.5 mmol). The solution was stirred for 8 to 16 h,then concentrated.

B. Part 2.

The residue from Part 2 (0.5 mmol) was dissolved in dichloromethane (3mL) and meta-chloroperbenzoic acid (2.5 mmol) was added. The solutionwas stirred for 8 to 16 h, the solvent was removed and residue purifiedby RP-HPLC to give Example 18 (4.3 mg). LCMS² Data: Molecular ion andretention time, 375.9 M+H); 3.37 min.

Example 195-Chloro-N-[1-(hydroxymethyl)-2,3-dimethylpentyl]thiophene-2-sulfonamide

A. Part 1

To a solution of sodium cyanide (735.15 mg, 15 mmol) and ammoniumcarbonate (1.92 g, 20 mmol) in EtOH/H₂O (1:1, 35 mL) was added 2,3dimethylpentanal (570.95 mg, 5 mmol). The solution was heated at 50° C.for 20 h, then concentrated.

B. Part 2

The residue from Part 1 (5 mmol) was dissolved in 35 mL of a 3N sodiumhydroxide solution and heated at 95° C. for 22 h. Stirring was continuedfor an additional 8 to 16 h, then the solvent was removed.

C. Part 3

To the residue from Part 2 (2.5 mmol) in THF (10 mL) was added lithiumaluminum hydride (1 M solution in THF) (5 mL, 5 mmol) and the solutionheated at 60° C. for 4 h. The solution was stirred at 25° C. for 8 to 16h. The reaction was quenched by addition of water (285 μL), 15% aqueoussodium hydroxide (285 μL), and water (665 μL) with vigorous stirringbetween each addition. The mixture was then filtered and concentrated.

D. Part 4

To the residue from Part 3 (0.5 mmol) in THF (5 mL) was addedtriethylamine (83.7 μL, 0.6 mmol) and 5-chlorothiophene-2-sulfonylchloride (108.54 mg, 0.5 mmol). The solution was stirred for 8 to 16 h,the solvent was removed and residue purified by RP-HPLC¹ to give Example19 (46.1 mg).

The following compounds (Examples 19-24, Table 5) were prepared using2,3 dimethylpentanal, 2-methylvaleraldehyde, 2-ethylhexanal,2,4,6-trimethyl-3-cyclohexene-1-carboxaldehyde,1,2,3,6-tetrahydro-benzaldehyde, cyclopentylmethanal, and following theprocedure outlined in Example 19. TABLE 5

(LCMS² Data: Molecular ion and retention time) R″SO₂Cl Aldehyde5-chlorothiophene-2-sulfonyl chloride 2,3 dimethylpentanal Example 19(326 M + H); 3.47 min 2-methylvaleraldehyde Example 20 (312 M + H); 3.25min 2-ethylhexanal Example 21 (340 M + H); 3.74 min2,4,6-trimethyl-3-cyclohexene-1- Example 22 carboxaldehyde (364 M + H);3.76 min 1,2,3,6-tetrahydrobenzaldehyde Example 23 (322 M + H); 3.11 minCyclopentylmethanal Example 24 (310 M + H); 3.07 min

Example 255-Bromo-N-[(1S)-1-(hydroxymethyl)-1,2-dimethylpropyl]thiophene-2-sulfonamide

A. Part 1

To a solution of (S)-α-methyl valine (131 mg, 1 mmol) in THF (5 mL) wasadded lithium aluminum hydride (1M solution in THF) (2 mL, 2 mmol) andthe solution was heated at 60° C. for 4 h. The solution was stirred at25° C. for 8 to 16 h. The reaction was quenched by addition of water(114 μL), 15% aqueous sodium hydroxide (114 μL), and water (266 μL) withvigorous stirring between each addition. The mixture was then filteredand concentrated.

B. Part 2

To the residue from Part 1 (0.5 mmol) in THF (2 mL) was addedtriethylamine (83.7 μL, 0.6 mmol) and 5-bromothiophene-2-sulfonylchloride (130.8 mg, 0.5 mmol). The solution was stirred for 8 to 16 h,the solvent was removed and residue purified by RP-HPLC¹ to give Example25 (50.8 mg).

The following compounds (Examples 25-26, Table 6) were prepared using5-bromothiophene-2-sulfonyl chloride and 5-chlorothiophene-2-sulfonylchloride and following the procedure outlined in Example 25. TABLE 6

(LCMS² Data: Molecular ion and retention time) RSO₂Cl S-α-methyl-valine5-bromothiophene-2-sulfonyl chloride Example 25 (344 M + H); 2.97 min5-chlorothiophene-2-sulfonyl chloride Example 26 (298 M + H); 2.92 min

Example 275-Chloro-N-[(1S,2R)-1-(hydroxymethyl)-2,4-dimethylpentyl]thiophene-2-sulfonamide

A. Part 1

A solution of 4-methyl-2-pentenoic acid (7.6 mL, 40 mmol) in THF (100mL) was cooled to −78° C. Triethylamine(5.85 mL, 42 mmol) andtrimethylacetyl chloride (pivaloyl chloride) (5.17 mL, 42 mmol) wereadded via syringe in that order. The dry ice lo bath was replaced withan ice bath and the reaction stirred at 0° C. for 1 h, then the reactionwas recooled to −78° C.

In a separate flask (R)-(+)-4-benzyl-2-oxazolidinone (7.0 g, 40 mmol)was dissolved in THF (100 mL) and cooled to −78° C., then n-butyllithium (1.6 M, 25 mL) was added via syringe. The mixture was stirredfor 20 min then the above reaction mixture added by removing the septumand pouring quickly from one flask to the other (Note: attempts totransfer reaction mixture via cannula failed due to the suspendedtrimethylammonium chloride in the mixture).

The resulting mixture was stirred at −78° C. for 30 min then allowed towarm to 25° C. for 1 to 2 h before quenching with saturated aqueousNH₄Cl solution (100 mL). Volatiles were removed on the rotary evaporatorand the aqueous slurry was diluted with water (200 mL) and extractedwith ethyl acetate (2×200 mL). The combined organic phase was dried overanhydrous MgSO₄, filtered and concentrated. The product may crystallizeout of solution and be of high purity. If purification is required thecrude product may be purified by flash chromatography using 20 to 30%ethyl acetate in hexane.

B. Part 2

To a copper (I) bromide/dimethyl sulfide complex (246 mg, 1.2 mmol) inTHF/DMS (2:1, 15 mL), cooled to −40° C., was added methyl magnesiumbromide (2.4 mL, 1 M solution in THF, 2.4 mmol). The solution wasallowed to stir for 10 min while warming to −15° C. The mixture wasrecooled to −40° C. and the product from Part 1 (245 mg, 1 mmol) in THF(6 mL) was added. The solution was stirred at 25° C. for 8 to 16 h. Thesolution was recooled to −78° C. and N-bromosuccinimide (356 mg, 2 mmol)in THF (2 mL) was added. The solution was allowed to warm to 0° C. andshaken at 0° C. for 3 h. The reaction was quenched with a 1:1 solutionof saturated ammonium carbonate and 0.5 N potassium bisulfate (5 mL).The organic phase was decanted off and concentrated.

C. Part 3

To the product from Part 3 dissolved in acetonitrile (5 mL) was addedtetramethylguanidine azide (0.6 mL, 4 mmol). The solution was stirredfor 72 to 120 h. The solution was concentrated to dryness, redissolvedin CH₂Cl₂ and 1 N HCl (2 mL) was added. The layers were separated andthe organic layer was filtered through a pad of silica gel washed withCH₂Cl₂(5 mL) and concentrated.

D. Part 4

To the product from Part 3 (131 mg, 1 mmol) in THF (5 mL) at 0° C. wasadded lithium aluminum hydride (1 M solution in THF) (2 mL, 2 mmol) andthe solution stirred at 25° C. for 4 h. The reaction was quenched byaddition of water (114 μL), 15% aqueous sodium hydroxide (114 μL), andwater (266 μL) with vigorous stirring between each addition. The mixturewas then filtered and concentrated.

E. Part 5

To the residue from Part 4 (0.5 mmol) in THF (2 mL) was addedtriethylamine (83.7 μL, 0.6 mmol) and 5-chlorothiophene-2-sulfonylchloride (108 mg, 0.5 mmol). The solution was stirred for 8 to 16 h, thesolvent was removed and residue purified as described for Example 1 togive 50.8 mg.

The following compounds (Examples 27-55, Table 7) were prepared using5-chlorothiophene-2-sulfonyl chloride with crotonic acid, 2-pentenoicacid, 2-hexenoic acid, 2-octenoic acid, cinnamic acid, furylacrylicacid, 4-methyl-2-pentenoic acid, and 4-phenylcinnamic acid and methyl,ethyl, isobutyl, 4-methoxyphenyl, hexyl and phenyl magnesium bromide andfollowing the procedure outlined in Example 27. TABLE 7

(LCMS² Data: Molecular ion and retention time) R′MgX 4- methoxy R methylethyl Isobutyl phenyl hexyl phenyl n-propyl methyl Ex. 27 Ex. 28 Ex. 29(326 (376 (354 M + H); M + H); M + H); 3.50 min 3.13 min 4.05 min ethylEx. 30 Ex. 31 Ex. 32 (312 (340 (390 M + H); M + H); M + H); 3.18 min3.69 min 3.32 min n-propyl Ex. 33 Ex. 34 Ex. 35 Ex. 36 Ex. 37 Ex. 38 Ex.39 (312 (326 (354 (404 (382 (374 (340 M + H); M + H); M + H); M + H);M + H); M + H); M + H); 3.26 min 3.49 min 3.93 min 3.66 min 4.46 min3.744 3.6 min min pentyl Ex.40 Ex. 41 Ex. 42 Ex. 43 Ex. 44 Ex. 45 (340(354 (382 (432 (410 (402 M + H); M + H); M + H); M + H); M + H); M + H);3.79 min 4.00 min 4.39 min 4.11 min 4.57 min 4.185 min phenyl Ex. 46 Ex.47 (346 (388 M + H); M + H); 3.27 min 3.91 min 2-furyl Ex. 48 Ex. 49 Ex.50 Ex. 51 (336 (350 (378 (406 M + H); M + H); M + H); M + H); 3.00 min3.28 min 3.69 min 4.19 min i-propyl Ex. 52 (382 4.47 min biphenyl Ex. 53Ex. 54 Ex. 55 (422 (436 (464 M + H); M + H); M + H); 3.90 min 4.14 min4.46 min

The following compounds (Examples 56-76, Table 8) were prepared using5-bromothiophene-2-sulfonyl chloride with crotonic acid, 2-pentenoicacid, 2-hexenoic acid, 2-octenoic acid, cinnamic acid,β-(3-pyridyl)-acrylic acid, furylacrylic acid, 4-methyl-2-pentenoicacid, and 4-phenylcinnamic acid and methyl, ethyl, isobutyl,4-methoxyphenyl, hexyl and phenyl magnesium bromide and following theprocedure outlined in Example 27. TABLE 8

(LCMS² Data: Molecular ion and retention time) R′MgX 4-methoxy R methylethyl isobutyl phenyl Hexyl methyl Ex. 56 (372 Ex. 57 (400 M + H); M +H); 3.52 min 4.07 min ethyl Ex. 58 (358 Ex. 59 (386 M + H); M + H); 3.26min 3.71 min n-propyl Ex. 60 (372 Ex. 61 (400 M + H); M + H); 3.52 min3.95 min pentyl Ex. 62 (400 Ex. 63 (428 Ex. 64 (478 Ex. 65 (456 M + H);M + H); M + H); M + H); 4.02 min 4.41 min 4.12 min 4.57 min phenyl Ex.66 (392 Ex. 67 (405 Ex. 68 (434 M + H); M + H); M + H); 3.31 min 3.55min 3.93 min pyridyl Ex. 69 (433 M + H); 2.67 min 2-furyl Ex. 70 (382Ex. 71(395 Ex. 72 (424 Ex. 73 (452 M + H); M + H); M + H); M + H); 3.04min 3.32 min 3.71 min 4.21 min i-propyl Ex. 74 (372 Ex. 75 (400 M + H);M + H); 3.49 min 3.96 min biphenyl Ex. 76 (482 M + H); 4.16 min

Example 77A5-Chloro-N-[(1S,2R)-2-ethyl-1-(hydroxymethyl)octyl]thiophene-2-sulfonamide

Following the procedure outlined in Example 27 (Part 1 and 2),2-pentenoic acid was coupled with 4R-4-benzyl-2-oxazolidinone to giveR-3-(2′-pentenyl)-4-benzyl-2-oxazolidinone. Addition of hexyl magnesiumbromide was followed by trapping by N-bromosuccinimide. After workup,flash chromatography over silica gel using 5% ether in hexane, gaveapproximately a 2:1 mixture of (1R-2R)-:(1R-2S)-3-(2′-bromo-3′ethylnonanyl)-4-benzyl-2-oxazolidinone.

Each isomer was converted to the corresponding sulfonylated aminoalcohol following the procedure in Example 27, (Steps 3-5). TABLE 9

(LCMS² Data: Molecular ion and retention time) 5-chlorothiophene-2-5-bromothiophene-2- sulfonyl sulfonyl 1S-2R Example 77A Example 78A (368M + H) (414 M + H) 4.24 min 4.26 min 1S-2S Example 77B Example 78B (368M + H) (414 M + H) 4.24 min 4.26 min

Example 795-Chloro-N-[(1S)-1-(hydroxymethyl)-2-(methylamino)butyl]thiophene-2-sulfonamide

A. Part 1:

To a solution of D-serine (1.05 g, 10 mmol) in H₂O/THF (1:1, 100 mL) at0° C. was added sodium hydroxide (2.17 g, 30 mmol) and5-chlorothiophene-2-sulfonyl chloride (2.17 g, 10 mmol). The solutionwas stirred for 2 to 3 h, then the organic phase was concentrated andthe aqueous phase acidified with 1 N HCl and extracted into ethylacetate and concentrated.

B. Part 2:

To the residue from Part 1 (2.5 mmol) dissolved in THF (25 mL) at −78°C. was added ethyl magnesium bromide (7.5 mL, 7.5 mmol). The mixture waswarmed to 25° C. and allowed to stir for 48 h. It was then acidifiedwith 1 N HCl and extracted into ethyl acetate and concentrated.

C. Part 3:

To the product from Part 2 (0.1 mmol) dissolved in DMF (500 μL) wasadded CH₂Cl₂ (1.5 mL), acetic acid (12 μL, 0.2 mmol) and methyl amine (2M solution in THF) (100 μL, 0.2 mmol). The reaction was stirred for 5min and sodium triacetoxyborohydride (105.6 mg, 0.5 mmol) was added. Thesolution was allowed to stir for 8-16 h and purified by RP-HPLC¹ to giveExample 79 (6.8 mg).

The following compounds (Examples 79-86, Table 10) were prepared usingmethyl, ethyl or pentyl magnesium bromide with methylamine (2M soln inTHF), ethylamine (2M soln in THF), ethanolamine, benzylamine, andcyclopentylamine and following the procedure outlined in Example 79.TABLE 10

(LCMS² Data: Molecular ion and retention time) R′MgBr NHR″ methyl ethylpentyl Methylamine Example 79 (313.0 M + H); 1.67 min ethylamine Example80 (313.0 M + H); 1.53 min Ethanolamine Example 81 Example 82 Example 83(329.0 M + H); (343.0 M + H); (385.0 M + H); 1.22 min 1.73 min 2.36 minBenzylamine Example 84 Example 85 (375.0 M + H); (389.0 M + H); 2.12 min2.25 min cyclopentylamine Example 86 (353.0 M + H); 1.99 min

Example 875-Chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-N-(2-phenoxyethyl)thiophene-2-sulfonamide

A. Part 1

To a solution of L-isoleucine methyl ester hydrochloride (1.82 g, 10mmol) and 5-chlorothiophene-2-sulfonyl chloride (1.82 g, 10 mmol) wasadded triethylamine (4.18 mL, 30 mmol). The mixture was stirred at 60°C. overnight, then filtered and concentrated. The crude product waspurified by flash chromatography over silica gel using 10% ethyl acetatein hexane to give 5-chlorothiophene-2-sulfonyl isoleucine methyl ester2.53 g.

B. Part 2

To a solution of 5-chlorothiophene-2-sulfonyl isoleucine methyl ester(103 mg, 0.25 mmol) in DMF (1 mL) was added β-bromophenetole (55 mg, 0.5mmol) and potassium carbonate (103 mg, 0.75 mmol. The reaction wasshaken at 25° C. overnight, then concentrated.

C. Part 3

The residue from part 2 was dissolved in 5% methanol in THF (1 mL) andlithium borohydride (11 mg, 0.5 mmol) added. The reaction was shaken at25° C. for 2 days then quenched by addition of water (1 mL) andextracted into ethyl acetate (3.5 mL). The organic phase was evaporatedand the residue purified by RP-HPLC¹ to give Example 87 (48 mg).

The following compounds (Examples 87-88, Table 11) were prepared usingβ-bromophenetole, 3-chlorobenzyl bromide and following the procedureoutlined in Example 87. TABLE 11 (LCMS² Data: Molecular ion andretention time)

RSO₂Cl R₁Br 5-chlorothiophene-2-sulfonyl chloride β-bromophenetoleExample 87 (418.0 M + H); 4.05 min 3-chlorobenzyl bromide Example 88(422.0 M + H); 4.12 min

Example 895-Chloro-N—[(S)-2-hydroxy-1-phenylethyl]thiophene-2-sulfonamide

To a solution of (S)-(+)-2-phenylglycinol (6.8 mg, 0.05 mmol) in CH₃CN(200 μL) was added Et₃N (105 μL, 1M in CH₃CN) and5-chlorothiophene-2-sulfonyl chloride (10.9 mg, 0.05 mmol) as a solutionin CH₃CN (200 μL). The vial was capped and shaken for 8 to 12 h at 40°C. Solvent was removed in vacuo, and the residue dissolved in 1.6 mLDMSO (0.03 M).

The following compounds (Examples 89-117, Table 12) were prepared using5-chlorothiophene-2-sulfonyl chloride and 5-bromothiophene-2-sulfonylchloride with (S)-(+)-2-phenylglycinol, L-leucinol,DL-2-amino-1-hexanol, 2-amino-2-methyl-1-propanol,2-amino-2-ethyl-1,3-propanediol, cycloleucinol, (S)-cyclohexylalaninol,L-phenylalaninol, L-methioninol, DL-2-amino-1-pentanol, L-tert-leucinol,chloramphenicol, (S)-(+)-2-amino-1-butanol, (S)-benzyl-L-cysteinol,benzyl-L-threoninol, 4-methylbenzyl-H-cysteinol, benzyl-H-tyrosinol, andL-threoninol following the procedure outlined in Example 89. TABLE 12(LCMS Data: Molecular ion and retention time) Amine X = Cl X = Br(S)-(+)-2-phenylglycinol Example 89 Example 104 (316.46 M-H), 0.95(361.31 M-H); 0.98 min min L-leucinol Example 90 Example 105 (296.48M-H), 1.01 (342.41 M-H); 1.02 min min DL-2-amino-1-hexanol Example 91Example 106 (296.49 M-H), 1.02 (342.39 M-H); 1.04 min min2-amino-2-methyl-1-propanol Example 92 Example 107 (268.45 M-H), 0.81(314.38 M-H); 0.83 min min 2-amino-2-ethyl-1,3- Example 93 Example 108propanediol (298.46 M-H), 0.69 (344.37 M-H); 0.69 min min cycloleucinolExample 94 Example 109 (295.02 M-H), 0.92 (340.4 M-H); 0.93 min min(S)-cyclohexylalaninol Example 95 Example 110 (336.31 M-H), 1.19 (382.41M-H); 1.2 min min L-phenylalaninol Example 96 (330.50 M-H), 1.03 minL-methioninol Example 111 (360.33 M-H); 0.9 min DL-2-amino-1-pentanolExample 97 (Example 112 (282.68 M-H), 0.92 327.07 M-H); 0.94 min minL-tert-leucinol Example 98 Example 113 (296.50 M-H), 1.22 (341.44 M-H);1.01 min min Chloramphenicol Example 99 (391.44 M-H), 0.89 min(S)-(+)-2-amino-1-butanol Example 100 (268.45 M-H), 0.61 minS-benzyl-L-cysteinol Example 101 Example 114 (377.77 M-H), 1.13 (422.35M-H); 1.15 min min benzyl-L-threoninol Example 102 (374.49 M-H), 1.21min 4-methylbenzyl-H-cysteinol Example 115 (436.31 M-H); 1.01 minbenzyl-H-tyrosinol Example 116 (480.43 M-H); 1.15 min L-threoninolExample 103 Example 117 284.10 (M-H), 0.58 329.99 (M-H), 0.68 min min

Example 1185-Chloro-N—[(S,S)-1-formyl-2-methylbutyl]thiophene-2-sulfonamide

A. Part 1

To a solution of 5-chlorothiophene-2-sulfonyl chloride (11 g, 50.7 mmol)in CH₃CN (100 mL) and (S)-isoleucinol (6.2 g, 53 mmol) was added Et₃N(11 mL, 109 mmol). The reaction mixture was heated at 50° C. withstirring for 24 h. The solvent was removed and the oil was dissolved inEtOAc (100 mL). The solution was washed with water (2×100 mL), brine(1×100 mL), and dried over Na₂SO₄. The solvent was removed to give 13.85g (88%) of the desired sulfonamide.

B. Part 2

Molecular sieves (15 g, 4 Å) were stirred in dry CH₂Cl₂ (175 mL) for 10min. Then a mixture of pyridinium chlorochromate (8.6 g, 39.9 mmol) andsilica gel (9 g) 15 was added and the mixture was stirred an additional10 min. To the suspension was added 5-chlorothiophene-2-sulfonylisoleucinol (4 g, 13.4 mmol) dissolved in CH₂C1₂ (15 mL) and theresulting slurry was stirred for 2 h. The reaction mixture was filteredand the solvent was removed. The residue was subjected to a Biotage™eluting with 20% EtOAc/hexane to give 3.22 g (81%) of the aldehyde(LCMS=294.21 (M−H), rt=1.10 min).

Example 1195-Chloro-N—[(S,S)-1-(1-hydroxyethyl)-2-methylbutyl]thiophene-2-sulfonamide

To a solution of the aldehyde from example 118 (23.7 mg, 0.08 mmol) inTHF (400 μL) was added methyl magnesium bromide (400 μL, 1.0 M in THF, 5eq). The vial was capped and agitated at 50° C. for 12 h. The reactionwas quenched with sat. aqueous NH₄Cl (1.5 mL) and EtOAc (1 mL). Theorganic layer was transferred into a tarred vial and the aqueous layerwas extracted with EtOAc (1 mL). The combined organics were concentrated(Savant, medium heat) and the resulting mixture of diastereomers wasdissolved in DMSO such that the final concentration was 360 mM.

The following compounds (Examples 119-154, Table 13) were prepared using5-chlorothiophene-2-sulfonyl isoleucinal (example 118) and5-bromothiophene-2-sulfonyl isoleucinal (prepared as in example 118)with methylmagnesium bromide, cyclopentylmagnesium bromide,hexylmagnesium bromide, pentylmagnesium bromide, butylmagnesium bromide,isopropylmagnesium bromide, o-tolylmagnesium bromide,tert-butylmagnesium bromide, isobutylmagnesium bromide, vinylmagnesiumbromide, allylmagnesium bromide, ethylmagnesium bromide,4-fluorophenylmagnesium bromide, 4-chlorophenylmagnesium bromide,2-methyl-1-propenylmagnesium bromide, isopropenylmagnesium bromide,4-anisylmagnesium bromide, 1-methyl-1-propenylmagnesium bromide,2-[2-(1,3-dioxanyl)]ethylmagnesium bromide, 3-butenylmagnesium bromide,1-propynylmagnesium bromide, 4-thioanisolemagnesium bromide, and4-N,N-dimethylanilinemagnesium bromide following the procedure outlinedin example 119. Note: during the reaction sequence with the5-bromothiophene compounds, the bromine is converted to a hydrogen onthe thiophene ring. TABLE 13 (LCMS Data: Molecular ion and retentiontime)

R—MgBr 5-chlorothiophene thiophene methylmagnesium bromide Ex. 119310.09(M − H), 1.06 min 310.10(M − H), 1.12 min cyclopentylmagnesiumbromide Ex. 120 Ex. 140 364.13(M − H), 1.41 min 330.19(M − H), 1.26 minHexylmagnesium bromide Ex. 121 Ex. 141 380.16(M − H), 1.50 min 346.24(M− H), 1.38 min 380.17(M − H), 1.54 min 346.24(M − H), 1.42 minpentylmagnesium bromide Ex. 122 Ex. 142 366.15(M − H), 1.42 min 332.19(M− H), 1.30 min 366.16(M − H), 1.47 min 332.19(M − H), 1.35 minButylmagnesium bromide Ex. 123 Ex. 143 352.15(M − H), 1.34 min 318.18(M− H), 1.26 min 352.13(M − H), 1.40 min isopropylmagnesium bromide Ex.124 338.11(M − H), 1.31 min o-tolylmagnesium bromide Ex. 144 352.16(M −H), 1.24 min tert-butylmagnesium bromide Ex. 125 Ex. 145 352.14(M − H),1.41 min 318.2(M − H), 1.28 min isobutylmagnesium bromide Ex. 126352.14(M − H), 1.33 min 352.13(M − H), 1.38 min vinylmagnesium bromideEx. 127 Ex. 146 322.09(M − H), 1.14 min 288.15(M − H), 0.98 min 322.10(M− H), 1.19 min 288.15(M − H), 1.02 min allylmagnesium bromide Ex. 128Ex. 147 336.11(M − H), 1.22 min 302.17(M − H), 1.06 min 336.12(M − H),1.27 min 302.17(M − H), 1.11 min ethylmagnesium bromide Ex. 129 Ex. 148324.10(M − H), 1.18 min 290.18(M − H), 1.01 min 324.11(M − H), 1.22 min290.17(M − H), 1.06 min 4-chlorophenylmagnesium bromide Ex. 131 406.06(M− H), 1.36 min 406.06(M − H), 1.41 min 2-methyl-1-propenylmagnesium Ex.132 bromide 350.13(M − H), 1.25 min 350.10(M − H), 1.31 minisopropenylmagnesium bromide Ex. 133 336.11(M − H), 1.25 min 336.10(M −H), 1.28 min 4-anisylmagnesium bromide Ex. 134 Ex. 149 402.13(M − H),1.25 min 368.16(M − H), 1.14 min 402.12(M − H), 1.31 min 368.16(M − H),1.16 min 1-methyl-1-propenylmagnesium Ex. 135 bromide 350.13(M − H),1.27 min 350.12(M − H), 1.35 min 2-[2-(1,3-dioxanyl)] Ex. 136 Ex. 150ethylmagnesium bromide 410.15(M − H), 1.15 min 376.19(M − H), 0.98 min3-butenylmagnesium bromide Ex. 137 Ex. 151 350.11(M − H), 1.30 min316.17(M − H), 1.15 min 350.12(M − H), 1.33 min 316.17(M − H), 1.18 min1-propynylmagnesium bromide Ex. 152 300.17(M − H), 1.20 min (M − H), min4-thioanisolemagnesium bromide Ex. 138 Ex. 153 418.11(M − H), 1.35 min384.13(M − H), 1.26 min 418.11(M − H), 1.39 min (M − H), min4-N,N-dimethylanilunemagriesium Ex. 139 Ex. 154 bromide 415.15(M − H),0.89 min 381.21(M − H), 0.68 min 415.17(M − H), 0.93 min 381.21(M − H),0.71 min

Example 1555-Chloro-N—{(S,S)-1-[(S)-cyclohex-2-en-1-yl(hydroxy)methyl]-2-methylbutyl}thiophene-2-sulfonamide

To a two dram vial containing magnesium turnings (60 mg, 2.5 mmol)suspended in THF (3 mL) was added 2-bromocyclohexene (288 μL, 2.5 mmol)followed by 5-chlorothiophene-2-sulfonyl isoleucinal (1 mL of a 1 M THFsolution, 1 mmol, example 118). The vial was capped and agitated at 50°C. for 18 h. The vial was cooled and sat. aqueous NH₄Cl (1 mL) wasadded. The vial was vortexed and the organic layer was transferred intoa tarred vial and the aqueous layer was extracted with EtOAc (1 mL). Thecombined organics were concentrated in vacuo and the residue wassubmitted to semipreparative RP-HPLC using the conditions below.

-   Semi-preparative RP-HPLC conditions.-   Column: Spring Axial compression; Kromasil C18 10 μm particle size;    50×150 mm-   Solvent A: Water (0.1% TFA)-   Solvent B: Acetonitrile-   Solvent Gradient: 15-95% over 24 min, full cycle is 35 min-   Flow Rate: 60ml/min-   The product peak was collected based on UV (or ELSD) absorption.

The following compounds (Examples 155-161, Table 14) were prepared using2-bromocyclohexene, crotyl bromide, 1-bromo-2-pentene,3-bromo-2-methylpropene, and cinnamyl bromide following the procedureoutlined in Example 155. TABLE 14 (LCMS Data: Molecular ion andretention time)

Alkyl bromide 2-bromocyclohexene Example 155 376.70(M − H), 1.27 mincrotyl bromide Example 156 350.50(M − H), 1.22 min crotyl bromideExample 157 350.70(M − H), 1.25 min 1-bromo-2-pentene Example 158364.60(M − H), 1.35 min 3-bromo-2- Example 159 methylpropene 350.60(M −H), 1.20 min 3-bromo-2- Example 160 methylpropene 350.60(M − H), 1.24min cinnamyl bromide Example 161 412.60(M − H), 1.34 min

Example 1625-Chloro-N—[(S,S)-1-(1-hydroxy-1-methylethyl)-2-methylbutyl]thiophene-2-sulfonamide

A. Part 1

To a solution of 5-chlorothiophene-2-sulfonyl chloride (1.09 g, 5 mmol)in CH₃CN (20 mL) was added (L)-isoleucine methyl ester hydrochloride(908.5 mg, 5 mmol) as a solution in CH₃CN (10 mL) and Et₃N (1 mL, 7.2mmol). The reaction mixture was heated at 50° C. with shaking for 3days. The solvent was removed and the oil was dissolved in EtOAc (10mL). The solution was washed with water (5 mL), sat. NH₄OH (5 mL), andbrine (5 mL), and dried over MgSO₄. The solvent was removed to give 1.44g (88%) of the desired sulfonamide.

B. Part 2

To a solution of the ester from part 1 (40.7 mg, 0.125 mmol) in THF (500μL) was added methyl magnesium bromide (333 μL, 3.0 M in THF, 8 eq). Thevial was capped and agitated at 50° C. for 12 h. The reaction wasquenched with sat. aqueous NH₄Cl (1.5 mL) and EtOAc (1 mL). The organiclayer was transferred into a tarred vial and the aqueous layer wasextracted with EtOAc (1 mL). The combined organics were concentrated(Savant, medium heat) and the product was dissolved in DMSO such thatthe final concentration was 30 mM.

The following compounds (Examples 162-176, Table 15) were prepared using5-chlorothiophene-2-sulfonyl isoleucine methyl ester and5-bromothiophene-2-sulfonyl isoleucine methyl ester (from part 2) withmethylmagnesium bromide, pentylmagnesium bromide, phenylmagnesiumbromide, allylmagnesium bromide, ethylmagnesium bromide,4-chlorophenylmagnesium bromide, isopropenylmagnesium bromide,4-anisylmagnesium bromide, 1-methyl-1-propenylmagnesium bromide,3-butenylmagnesium bromide, 1-propynylmagnesium bromide,1-naphthylmagnesium bromide following the procedure outlined in example162. TABLE 15 (LCMS Data: Molecular ion and retention time)

R—MgBr X = Cl X = Br methylmagnesium bromide Example 162 324.58(M − H),1.19 min pentylmagnesium bromide Example 163 436.69(M − H), 1.79 minphenylmagnesium bromide Example 164 448.60(M − H), 1.50 minAllylmagnesium bromide Example 165 376.58(M − H), 1.44 minethylmagnesium bromide Example 166 Example 173 352.63(M − H), 1.39395.49(M − H), 1.45 min min 4-chlorophenylmagnesium bromide Example 167516.49(M − H), 1.62 min isopropenylmagnesium bromide Example 168 Example174 376.60(M − H), 1.51 421.29(M − H), 1.39 min min 4-anisylmagnesiumbromide Example 169 508.59(M − H), 1.45 min 1-methyl-1-propenylmagnesiumExample 170 Example 175 bromide 404.61(M − H), 1.65 447.46(M − H), 1.52min min 3-butenylmagnesium bromide Example 171 Example 176 404.64(M −H), 1.54 447.25(M − H), 1.26 min min 1-naphthylmagnesium bromide Example172 548.56(M − H), 1.64 min

Example 1775-Chloro-N-[1-(hydroxymethyl)cyclohexyl]thiophene-2-sulfonamide

A. Part 1

To a suspension of 1-amino-1-cyclohexane carboxylic acid (5 g, 35 mmol)and THF (100 mL) was added borane dimethyl sulfide (50 mL, 2M in THF) at0° C. The cold bath was allowed to expire and the reaction was stirredat 25° C. overnight. NaOH (3M, 100 mL) was added and the mixture wasstirred for 4 h. The reaction mixture was saturated with K₂CO₃ andextracted with Et₂O (2×100 mL). The combined organics were washed withbrine (100 mL) and dried over MgSO₄ to give 4.35 g (96%) of the desiredamino alcohol.

B. Part 2

The amino alcohol was sulfonylated as in Example 89.

The following compounds (Examples 177-183, Table 16) were prepared usingthe amino alcohols of 1-amino-1-cyclohexane carboxylic acid,2-amino-2-norbornane carboxylic acid, D,L-1-aminoindane-1-carboxylicacid, and 2-aminoindane-2-carboxylic acid hydrochloride with5-chlrorothiophene-2-sulfonyl chloride and 5-bromothiophene-2-sulfonylchloride following the procedure outlined for Example 177. TABLE 16(LCMS Data: Molecular ion and retention time)

Aminoacid X = Cl X = Br 1-amino-1-cyclohexane carboxylic Example 177Example 181 acid 308.14(M − H), 1.00 min 353.99(M − H), 1.02 min2-amino-2-norbomane carboxylic Example 178 Example 182 acid 320.13(M −H), 1.04 min 366.03(M − H), 1.06 min D,L-1-aminoindane-1-carboxylicExample 179 acid 342.12(M − H), 1.09 min 2-aminoindane-2-carboxylic acidExample 180 Example 183 HCl 342.12(M − H), 1.07 min 388.01(M − H), 1.08min

The following compounds (Examples 184-195, Table 17) were synthesizedusing 5-chlorothiophene-2-sulfonyl isoleucinal and5-bromothiophene-2-sulfonyl isoleucinal with methylmagnesium bromide,n-propylmagnesium chloride, and allylmagnesium bromide following theprocedure outlined for example 119. The resulting mixtures ofdiastereomers were isolated by semi-preparative RP-HPLC using theconditions outlined for example 155. TABLE 17 (LCMS Data³: Molecular ionand retention time) R-MgX X = Cl X = Br methylmagnesium Example 184Example 190 bromide 310.41 (M-H), 0.87 356.20 (M-H), 0.95 min minExample 185 Example 191 310.44 (M-H), 0.93 356.15 (M-H), 1.02 min minn-propylmagnesium Example 186 Example 192 chloride 338.47 (M-H), 1.06384.17 (M-H), 1.13 min min Example 187 Example 193 338.45 (M-H), 1.12384.24 (M-H), 1.18 min min allylmagnesium bromide Example 188 Example194 336.10 (M-H), 1.2 382.00 (M-H), 1.2 min min Example 189 Example 195336.10 (M-H), 1.24 382.00 (M-H), 1.25 min min

The pure synthetic diastereomer of Example 189 was prepared as follows.

A. Part 1

To a −78° C. solution of the Weinreb amide (see: F. Roux, et. Al.Tetrahedron, 1994, 50 (18), 5345-5360) of BOC-protected isoleucine(13.17 g, 48 mmol) was added allylmagnesium bromide (90 mL, 1M in THF).The cold bath was allowed to expire and the reaction stirred at 25° C.overnight. The reaction was quenched by the addition of cold aq. HCl(150 mL, 1M). After 30 min of stirring, the layers were separated andthe aqueous layer extracted with ethyl acetate (3×75 mL). The combinedorganics were dried over MgSO₄ and the solvent removed to give 8.41 g(69%) of the desired ketone.

B. Part 2

To a solution of the ketone from part 1 (8.4 g) in MeOH (200 mL) wasadded NaBH₄ (1.5 g, 39.6 mmol) as a solid. The reaction was stirred at25° C. for 5 h at which time the solvent was removed in vacuo. Theresidue was dissolved in ethyl acetate (100 mL) and washed with water(2×50 mL). The crude product was subjected to a Biotage™ eluting with 5to 15% EtOAc/hexane to give 4.12 g (49%) of the desired alcohol.

C. Part 3

A solution of alcohol from part 2 (4.12 g, 16 mmol), CH₂Cl₂ (75 mL), andTFA (15 mL) was stirred at 25° C. for 15 min. The reaction was quenchedwith a solution of NaOH (15 mL, 1 M) and then basified to pH 12 withNaOH pellets. The resulting solution was extracted with CH₂Cl₂ (2×50mL), and the combined organics were washed with water (25 mL), brine (25mL), and dried over MgSO₄ to give 2.44 g (97%) of the desired aminoalcohol, which was carried on to part 4 without further purification.

D. Part 4

The amino alcohol was sulfonylated as in example 89. The pure syntheticdiastereomer of Example 193 was obtained as follows:

To a solution of the BOC-amino homoallyl alcohol (1.1 g, 4.27 mmol, seepart 1-2 of example 188) in absolute EtOH (50 mL) was added Pd/C (110mg). The flask was placed under an atmosphere of hydrogen (balloon) andstirred at 25° C. Following completion of the reaction (2 h), themixture was filtered through a pad of Celite and the solvent was removedto give 1.16 g (quant) of the propyl analog. The BOC group was removedand the amine sulfonylated per the protocol outlined-for example 189.

Example 1965-Chloro-N—[(S,S)-2-methyl-1-(2,2,2-trifluoro-1-hydroxyethyl)butyl]thiophene-2-sulfonamide

To a 0° C. solution of 5-chlorothiophene-2-sulfonyl isoleucinal (770 mg,2.6 mmol, see Example 118, parts 1&2) in THF (5 mL) was added TMS-CF₃ (5mL, 0.5M in THF). The resulting mixture was treated with TBAF (250 μL,1M in THF). The cold bath was removed and the reaction was stirred at25° C. overnight. The reaction was quenched with HCl (25 mL, 2M) and theresulting solution was extracted with ethyl acetate (3×15 mL). Thecombined organic extracts were washed with water (25 mL) and brine (25mL) then dried over MgSO₄. The residue was submitted to RP-HPLC (seeexample 155 for procedure) to give 74 mg of the desired product(m/z=364.0 (M−H), rt=1.23 min).

The following compounds (Examples 197-198, Table 18) were synthesizedusing 1-amino-1-cyclohexane carboxylic acid and5-chlorothiophene-2-sulfonyl chloride with allylmagnesium bromide and2-methyl allylmagnesium chloride following the four step procedureoutlined for examples 177 (parts 1&2), 118 (part 2), and 119respectively. TABLE 18 (LCMS Data: Molecular ion and retention time)

R—MgX allylmagnesium bromide Example 197 348.10(M − H), 1.18 min2-methylallylmagnesium Example 198 chloride 364.10(M − H), 1.26 min

Example 199A5-Chloro-N—[(S)-2-hydroxy-1-(4-hydroxycyclohexyl)ethyl]thiophene-2-sulfonamide

A. Part 1

To a solution of 4-hydroxy-L-phenylglycine (10 g, 60 mmole) in NaOH (20mL, 3M) was added water (380 mL) and Raney nickel (30 g). The reactionmixture was hydrogenated at about 3 atm at 60 to 80° C. for 36 h in ahydrogen bomb. The reaction mixture was filtered through Celite andreduced in volume to about 80-100 mL and dioxane (100 mL) was added. Theresulting mixture was cooled to 0° C. and treated with Et₃N (10 mL) and5-chlorothiophene-2-sulfonyl chloride (16 g, 72 mmoles). The reactionwas allowed to warm up to 25° C. and stirred overnight. The dioxane andEt₃N were removed and the remaining aqueous solution diluted with 1N aq.HCl. The resulting precipitate was collected, washed with water anddiethyl ether to give the desired product as a white solid (12 g, 50% intwo steps) (100% purity by ELSD, m/z=352 (M−1)).

B. Part 2

To a suspension of(S)—N-(5-Cl-thiophene-2-sulfonyl)-4-hydroxycyclohexylglycine (12 g,33.99 mmol, part 1) in anhydrous THF was added borane-THF (110 mL, 1 Min THF, 110 mmoles) dropwise at 0° C. The resulting mixture was stirredat 25° C. over the weekend. The reaction mixture was quenched with HCl(75 mL, 1M) at 0° C. and stirred at 25° C. for 1 h. THF was removed andthe precipitate was collected, washed with water (containing a smallamount of diethyl ether), and dried to give a white solid as the desiredproduct (9 g, 78%) (100% purity by ELSD, m/z=338.5 (M−1), HPLC retentiontime³=0.64 min).

Example 1995-Chloro-N—[(S)-2-hydroxy-1-(4-methoxycyclohexyl)ethyl]thiophene-2-sulfonamide

A. Part 1

A mixture of5-chloro-N—[(S)-2-hydroxy-1-(4-hydroxycyclohexyl)ethyl]thiophene-2-sulfonamide(6.4 g, 18.83 mmol) from Example 199A, 2,2-dimethoxypropane (7 mL, 5.65mmol), and TsOH H₂O (72 mg, 0.38 mmol.) in anhydrous benzene (120 mL)was refluxed. After one hour, benzene was slowly distilled underatmospheric pressure to a final volume of 10 mL. Fresh benzene (100 mL)and 2,2-dimethoxypropane (5 mL) were added and the above operation wasrepeated. The residue was partitioned between diethyl ether and sat.NaHCO₃. The aqueous layer was extracted with diethyl ether (3×100 mL),and the combined extracts were dried over MgSO₄. The crude product waspurified by column chromatography using 1:5 EtOAc/CH₂Cl₂ as an eluent togive the N,O-acetonide (5.77 g, 81%) (100%, m/z=380 (M+1)).

B. Part 2

To a 0° C. solution of(S)-2-(5-Cl-thiophene-2-sulfonamido)-2-(4-hydroxycyclohexyl)-N,O-acetonide(379 mg, 1 mmol) in THF (7 mL) and DMF (2 mL) was added NaH (80 mg, 2mmol). The resulting reaction was stirred at 0° C. for 10 min at whichtime iodomethane (311 μL, 5 mmol) was added. The reaction was allowed towarm to 25° C. and stirred for 18 h. The solvent was removed and aceticacid (80%, 15 mL) was added. The mixture was stirred at 25° C. over theweekend. Following removal of the acetic acid in vacuo, the residue wassubjected to column chromatography on silica gel eluting withMeOH/CH₂Cl₂ (3:10) to give 303 mg (86%) of the desired product.

The following compounds (Examples 199-202B, Table 19) were preparedusing(S)-2-(5-Cl-thiophenesulfonamido)-2-(4-hydroxycylohexyl)-N,O-acetonide(from Example 199, part 1) with iodomethane, 1-bromopropane, allylbromide, benzyl bromide, 2-picolyl chloride hydrochloride and 3-picolylchloride hydrochloride as outlined in Example 199. TABLE 19 (LCMS Data³:Molecular ion and retention time)

R—X iodomethane Example 199 352.1(M − H), 0.82 min 1-bromopropaneExample 200 366.0(M − H), 0.91 min allyl bromide Example 201 378.0(M −H), 1.01 min benzyl bromide Example 202 428.1(M − H), 1.21 min 2-picolylchloride HCl Example 202A 429.4(M − H), 0.57 min 3-picolyl chloride HClExample 202B 429.0(M − H), 0.57 min

Example 203N-[1-Acetyl-4-(hydroxymethyl)piperidin-4-yl]-5-chlorothiophene-2-sulfonamide

A. N-[1-Boc-4-(Carboxylicacid)piperidin-4-yl]-5-chlorothiophene-sulfonamide

Triethylamine (2.28 mL, 1.66 g, 16.45 mmol) was added to a slurry of1-Boc-4-aminopiperidine-4-carboxylic acid (2.68 g, 10.973 mmole) inacetonitrile:water (1:1) (40 mL) at 25° C. The slurry became a neonyellow to greenish solution at the end of addition. The slurry wasslightly warmed up (5 min) in order to obtain a solution. The mixturewas cooled to 0° C., 5-chlorothiophene-2-sulphonyl chloride (2.62 g,12.07 mmol) was added (5 min) dropwise as a solution in acetonitrile (8mL). The solution was allowed to warm up to 25° C. overnight. After 19h, an aliquot was taken. TLC (9:1 CH₂Cl₂:CH₃OH) indicated that reactionwas about 90% done. The reaction was quenched by addition of water (50mL), CH₂Cl₂ (50 mL) and ice cold 1N HCl (10 mL). The organic layer waswashed with water and saturated NaCl. It was dried over MgSO₄, filtered,and concentrated to a yellow oil (2.1 g). The crude material waspurified by column chromatography, silica gel 230 to 400 mesh, eluent:starting with 5% MeOH in CH₂Cl₂ and ending with 10% MeOH in CH₂Cl₂ tofurnish N-[1-Boc-4-(carboxylicacid)piperidin-4-yl]-5-chlorothiophene-2-sulfonamide as a whiteamorphous solid (1.2 g, 25.7%). Mass Spectrum (−ESI): 423 (M−H)⁻.

B.N-[1-Boc-4-(hydroxymethyl)piperidin-4-yl]-5-chlorothiophene-2-sulfonamide

1N Borane-THF (1.019 g, 12.14 mL, 11.86 mmol) was added dropwise over 30min at 0° C. to a solution of N-[1-Boc-4-(carboxylicacid)piperidin-4-yl]-5-chlorothiophene-2-sulfonamide (1.2 g, 2.82 mmol)in anhydrous tetrahydrofuran (15 mL). The reaction was allowed to warmup to 25° C. overnight, and then was quenched by addition of 30 mL of10% acetic acid in methanol. After solvent evaporation, the crudeproduct was dissolved in ethyl acetate and washed with 1M HCl, water and10% NaHCO₃. The organic layer was dried over MgSO₄, filtered andconcentrated to obtain a crude yellow oil (1.1 g). The crude product waspurified by column chromatography, silica gel 230-400 mesh, eluent:starting with 1:3 EtOAc-hexane and ending with 1:1 EtOAc-hexane toaffordN-[1-Boc-4-(hydroxymethyl)piperidin-4-yl]-5-chlorothiophene-2-sulfonamideas a colorless oil (0.79 g, 68.2%). Mass Spectrum (−ESI): 409 (M−H)⁻.

C. N-[4-(Hydroxymethyl)piperidin-4-yl]-5-chlorothiophene-2-sulfonamideHCl Salt

4N HCl (5 mL) was added to a stirred solution ofN-[1-Boc-4-(hydroxymethyl)piperidin-4-yl]-5-chlorothiophene-2-sulfonamide5 (0.7 g, 1.7 mmol) in EtOAc (4 mL). The solution was allowed to stir at25° C. After 30 min, a cloudy solution formed. After 2 h a precipitateformed. TLC (1: 1 EtOAc-hexane) indicated the reaction was complete. Thesolvent was reduced to ˜2-3 mL, diluted with diethyl ether (6 mL) andfiltered through a filter funnel. The precipitate was washed withdiethyl ether (3×5 mL) to obtainN-[4-(hydroxymethyl)piperidin-4-yl]-5-chlorothiophene-2-sulfonamide asan amorphous white solid (0.48 g, 90.7%). Mass Spectrum (+ESI): 311(M+H)⁺.

D.N-[1-Acetyl-4-(hydroxymethyl)piperidin-4-yl]-5-chlorothiophene-2-sulfonamide

Acetyl chloride (0.15 g, 1.894 mmol) was added dropwise (5 min) as asolution in CH₂Cl₂ (1 mL) to a cold 0° C. solution ofN-[4-(hydroxymethyl)piperidin-4-yl]-5-chloro-thiophene-2-sulfonamide(0.19 g, 0.61 mmol) in CH₂Cl₂ (5 mL) and triethylamine (0.44 mL, 3.18mmol) The solution was allowed to warm to 25° C. overnight (19 h). Analiquot was taken and TLC (1:1 EtOAc-hexane) indicated that reaction wascomplete. It was diluted with CH₂Cl₂ (10 mL) and the organic layer waswashed with 1N HCl (50 mL), saturated aqueous NaHCO₃ (50 mL) and NaCl(50 mL). The organic layer was dried over MgSO₄, filtered, andconcentrated to obtain a crude oil (175 mg). The crude product waspurified by column chromatography, silica gel 230-400 mesh, eluent:starting with 1:4 EtOAc-hexane and ending with 1:1 EtOAc-hexane toaffordN-[1-Acetyl-4-(hydroxymethyl)-piperidin-4-yl]-5-chlorothiophene-2-sulfonamideas yellowish color oil (62 mg, 28.9%). Mass Spectrum (+ESI): 353 (M+H)⁺.Anal. Calc'd for C₁₂H₁₇ClN₂O₄S₂.1.62 H₂O: C, 37.29; H, 5.70; N, 7.26.Found: C, 37.62; H, 5.36; N, 7.31.

Example 2045-Chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-furansulfonamide

A. 2-Chlorofuran

1.6 M nBuLi (15.37 g, 150 mL, 0.24 mol) was added dropwise over 10 minat 25° C. to a solution of furan (13.6 g, 0.20 mol) in dry diethyl ether(200 mL). When the dropwise addition was completed, the reaction mixturewas cooled to −70° C. At this temperature a solution of hexachloroethane(49.8 g, 0.21 mol) was added over 10 min and the temperature was notallowed to rise above −55° C. The reaction mixture was kept at −70° C.for 3 h. The reaction mixture was then warmed to 25° C., hydrolyzed withice water and neutralized with 2.5 N hydrochloric acid. The phases wereseparated and the water phase extracted twice with diethyl ether (100mL). The combined diethyl ether phases were washed once with a solutionof NaHCO₃ (50 mL) and once with water (50 mL) and dried over MgSO₄. Thediethyl ether was distilled off through a fractionating column and theproduct was collected at 78 to 79° C. to obtain 2-chlorofuran as acolorless oil (20.0 g, 97.6%). ¹H-NMR (DMSO-d₆, 400 MHz) δ 7.34 (d, 1H);6.38 (d, 1 H); 6.21 (d, 1H).

B. 5-Chlorofuran-2-sulfonyl chloride

Phosphorus pentachloride (40.53 g, 0.1947 mol) was added portionwise(caution, foaming) over 5 min at 25° C. to chlorosulfonic acid (56.8 g,32.4 mL, 0.487 mol) and the resulting solution was stirred at 25° C. for10 min. Then, 2-chlorofuran (20.0 g, 20 0.1947 mol) was added in oneportion and the resulting dark suspension was heated to 55° C. for 1.0 hduring which time foaming occurred and subsided. The reaction mixturewas then poured onto ice and the resulting suspension was extracted withCH₂Cl₂ (250 mL). The organic was filtered through a pad of celite,washed with brine (70 mL) and dried over MgSO₄. The solvent was removedin vacuo to provide 5-chlorofuran-2-sulfonyl chloride as a black oil(14.1 g, 36.02%). ¹H-NMR (DMSO-d₆, 400 MHz) δ 7.05 (d, 1H); 6.35 (d, 1H).

C.5-Chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-furansulfonamide

5-Chlorofuran-2-sulfonyl chloride (3.376 g, 16.79 mmol) was addeddropwise (5 min) as a solution in CH₂Cl₂ (10 mL) to a 0° C. solution ofL-isoleucinol (1.5 g, 12.92 mmol) in CH₂Cl₂ (15 mL) and triethylamine(2.69 mL, 19.38 mmol). The solution was allowed to warm to 25° C.overnight (19 h). An aliquot was taken and TLC (1:1 EtOAc-hexane)indicated that reaction was complete. It was diluted with CH₂Cl₂ (100mL) and the organic layer was washed with 1N HCl (2×50 mL), saturatedaqueous NaCl (50 mL). The organic layer was dried over MgSO₄, filtered,and concentrated to obtain a crude black oil (2.69 g). The crude productwas purified by column chromatography, silica gel 230 to 400 mesh,eluent: starting with 1:4 EtOAc-hexane and ending with 1:1 EtOAc-hexaneto afford5-chloro-N-[(1S,2S-1-(hydroxymethyl)-2-methylbutyl]-2-furansulfonamideas an amorphous white solid (0.98 g, 26.92%). Mass Spectrum (−ESI): 280(M−H)⁻. Anal. Calc'd for C₁₀H₁₆ClN₂O₄S: C, 42.63; H, 5.72; N, 4.97.Found: C, 42.34; H, 5.65; N, 4.77.

Example 205N-[(1S)-2-Butyl-1-(hydroxymethyl)hexyl]-5-chloro-2-thiophenesulfonamide

A. (4R)-4-Benzyl-3-[(E)-2-heptenoyl]-1,3-oxazolidin-2-one

Triethylamine (6.85 g, 49.15 mmol) and trimethyl acetyl chloride (6.05mL, 49.15 mmol) were added dropwise (5 min) to a −78° C. solution of2-heptenoic acid (6 g, 46.81 mmole) in THF (80 mL). The slurry wasstirred at −78° C. for 5 min and then replaced with a 0° C. coolingsystem. It was stirred at this temperature for 1 h. In a separate flask,a solution of R-(+)-4-benzyl-2-oxazolidinone (8.295 g, 46.81 mmol) wascooled to −78° C. and nBuLi (1.6M, 46.8 mmol) was added dropwise over 10min. The colorless solution was stirred at this temperature for 45 minand transferred via cannula to a −78° C. solution of the ester. Theyellowish slurry was warmed to 25° C. overnight (19 h). An aliquot wastaken and TLC (1:1 EtOAc-hexane) indicated that reaction was complete.It was cooled to 0° C. and the reaction was quenched by addition of H₂O(20 mL). It was diluted with ethyl acetate (200 mL) and the organiclayer was separated. The organic layer was dried over MgSO₄, filtered,and concentrated to obtain a crude yellow oil (13.69 g). The crudeproduct was purified by column chromatography, silica gel 230 to 400mesh, eluent: 1:4 EtOAc-hexane to obtain(4R)-4-benzyl-3-[(E)-2-heptenoyl]-1,3-oxazolidin-2-one as a colorlessoil (12.1 g, 92.80%). Mass Spectrum (−ESI): 288 (M−H)⁻.

B. (4R)-4-Benzyl-3-[(2R)-2-bromo-3-butylheptanoyl]-1,3-oxazolidin-2-one

A slurry of copper bromide (I) dimethyl sulfide complex (5.132 g, 24.967mmol) in THF (60 mL) and dimethyl sulfide (30 mL) as a co-solvent wascooled to −40° C. and n-butyl magnesium chloride (25 mL, 49.93 mmol) wasadded dropwise for 10 min and stirred for 20 min while warming to −15°C. The black slurry was cooled to -40° C. and(4R)-4-benzyl-3-[(E)-2-heptenoyl]-1,3-oxazolidin-2-one (6 g, 20.80 mmol)was added dropwise over 10 min as a solution in THF (20 mL) at −40° C.The reaction was let warm up to 25° C. overnight (20 h).N-Bromosuccinimide (7.407 g, 41.61 mmol) was added portionwise to a cold−78° C. solution of the black slurry. It was allowed to warm to 0° C.and was stirred for an additional 3 h. The reaction was quenched with a1:1 solution of saturated ammonium carbonate and 0.5 N potassiumbisulfate. The black slurry became greenish to blue. A precipitateformed (light blue). It was filtered. The mother liquor was diluted withethyl acetate (150 mL) and the organic was dried over MgSO₄, filtered,concentrated to obtain(4R)-4-benzyl-3-[(2R)-2-bromo-3-butylheptanoyl]-1,3-oxazolidin-2-one asa crude semi-solid (green) (8.49 g, 96.15%). Mass Spectrum (−ESI): 423(M−H)⁻.

C. (4R)-3-[(2S)-2-Azido-3-butylheptanoyl]-4-benzyl-1,3-oxazolidin-2-one

Tetramethylguanidine azide (TMGA) (5.398 g, 37.70 mmol) was addeddropwise (5 min) to a 25° C. solution of(4R)-4-benzyl-3-[(2R)-2-bromo-3-butylheptanoyl]-1,3-oxazolidin-2-one(4.0 g, 9.42 mmol) in acetonitrile (50 mL). The reaction was stirred for4 days. An aliquot was taken and TLC (1:4 EtOAc-hexane) indicated thatreaction was complete. The solvent was removed in vacuo. The resultingblack semi-solid was dissolved in CH₂Cl₂ (200 mL) and quenched with 1NHCl (30 mL). The organic layer was dried over MgSO₄, filtered, andconcentrated in vacuo to obtain(4R)-3-[(2S)-2-azido-3-butylheptanoyl]-4-benzyl-1,3-oxazolidin-2-one asa crude yellow oil (3.61 g, 99.1%). Mass Spectrum (−ESI): 385 (M−H)⁻.

D. (2S)-2-Amino-3-butyl-1-heptanol

To a slurry of LAH (1.219 g, 32.13 mmol) in THF (60 mL) was added(4R)-3-[(2S)-2-azido-3-butylheptanoyl]-4-benzyl-1,3-oxazolidin-2-one(3.6 g, 9.37 mmol) dropwise at 0° C. over 20 min. The reaction washeated to 36° C. for 18 h. The reaction slurry (brown) was cooled to 0°C. and the reaction was quenched with H₂O (15 mL) and washed with 1NNaOH (30 mL) and H₂O (15 mL). It was let stir for 2 h to obtain anoff-white slurry. The slurry was filtered and the mother liquor wasfurther dried over MgSO₄, filtered, and concentrated in vacuo to obtain(2S)-2-amino-3-butyl-1-heptanol as a crude yellow oil (1.93 g, 73.75%).Mass Spectrum (+ESI): 188 (M+H)⁺.

E.N-[(1S)-2-Butyl-1-(hydroxymethyl)hexyl]-5-chloro-2-thiophenesulfonamide

5-Chlorothiophene-2-sulfonyl chloride (2.42 g, 11.55 mmol) was addeddropwise (5 min) as a solution in CH₂Cl₂ (20 mL) to a 0° C. solution of(4R)-3-[(2S)-2-azido-3-butylheptanoyl]-4-benzyl-1,3-oxazolidin-one (1.9g, 10.14 mmol) and triethylamine (2.11 mL, 15.21 mmol). The solution wasallowed to warm to 25° C. overnight (19 h). An aliquot was taken and TLC(1:1 EtOAc-hexane) indicated that reaction was complete. It was dilutedwith CH₂Cl₂ (100 mL) and the organic layer was washed with IN HCl (2×50mL) and saturated aqueous NaCl (50 mL). The organic layer was dried overMgSO₄, filtered, and concentrated to obtain a crude oil (2.98 g). Thecrude product was purified by column chromatography, silica gel 230-400mesh, eluent: starting with 1:3 EtOAc-hexane and ending with 1:2EtOAc-hexane to furnishN-[(1S)-2-butyl-1-(hydroxymethyl)hexyl]-5-chloro-2-thiophenesulfonamideas an amorphous white solid (0.630 g, 16.9%). Mass Spectrum (−ESI): 366(M−H)⁻. Anal. Calc'd for C₁₅ H₂₅ N Cl O₃ S₂: C, 48.96; H, 7.12; N, 3.81.Found: C, 49.08, H, 6.83, N, 3.82.

Example 206N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-furansulfonamide

A. 2-Furansulfonyl chloride

Phosphorus pentachloride (15.29 g, 73.44 mmol) was added portionwise(caution, foaming) over 5 min at 0° C. to chlorosulfonic acid (21.39 g,183.6 mmol) and the resulting solution was stirred at 0° C. for 10 min.Then, furan (5.0 g, 73.44 mmol) was added in one portion and theresulting dark suspension was stirred at 0° C. for 15 min during whichtime foaming occurred and subsided. The reaction mixture was then pouredonto ice and the resulting suspension was extracted with CH₂Cl₂ (150mL). The organic extract was filtered through a pad of celite, washedwith brine and dried over MgSO₄. The solvent was removed in vacuo toprovide 2-furansulfonyl chloride as a black oil (1.01 g, 7.9%). ¹H-NMR(DMSO-d₆, 400 MHz) δ 7.4 (d, 1H); 6.38 (d, 1H); 6.35 (d, 1 H).

B. N-[(1S,2S)-1-(Hydroxymethyl)-2-methylbutyl]-2-furansulfonamide

2-Furansulfonyl chloride (1.01 g, 8.69 mmol) was added dropwise (5 min)as a solution in CH₂Cl₂ (5 mL) to a 0° C. solution of L-isoleucinol(0.909 g, 7.83 mmol) in CH₂Cl₂ (20 mL) and triethylamine (2.42 mL, 17.38mmol). The solution was allowed to warm to 25° C. overnight (19 h). Analiquot was taken and TLC (1:1 EtOAc-hexane) indicated that reaction wascomplete. It was diluted with CH₂Cl₂ (100 mL) and the organic layer waswashed with 1N HCl (2×50 mL) and saturated aqueous NaCl (50 mL). Theorganic layer was dried over MgSO₄, filtered, and concentrated to obtaina crude black oil (0.65 g). The crude product was purified by columnchromatography, silica gel 230-400 mesh, eluent: starting with 1:3EtOAc-hexane and ending with 1:2 EtOAc-hexane to furnishN-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-furansulfonamide as anamorphous white solid (0.155 g, 72.12%)

Mass Spectrum (−ESI): 246 (M−H)⁻. Anal. Calc'd for C₁₀H₁₇ClNO₄S: C,48.57; H, 6.93; N, 5.66. Found: C, 48.72; H, 6.78; N, 5.39.

Example 207N-[(1S,2S)-1-(Hydroxymethyl)-2-methylbutyl]-5-iodo-2-thiophenesulfonamide

A.5-Bromo-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-thiophenesulfonamide

5-Bromothiophene-2-sulfonyl chloride (5.0 g, 19.11 mmol) was addeddropwise (5 min) as a solution in CH₂Cl₂ (10 mL) to a 0° C. solution ofL-isoleucinol (2.108 g, 18.16 mmol) in CH₂Cl₂ (15 mL) and triethylamine(3.77 mL, 27.24 mmol). The solution was allowed to warm to 25° C.overnight (19 h). An aliquot was taken and TLC (1:1 EtOAc-hexane)indicated that reaction was complete. It was diluted in methylenechloride (100 mL) and the organic layer was washed with 1N HCl (2×50 mL)and saturated aqueous NaCl (50 mL). The organic layer was dried overMgSO₄, filtered, and concentrated to obtain a crude off-yellow solid(5.2 g). The crude product was purified by column chromatography, silicagel 230-400 mesh, eluent: starting with 1:4 EtOAc-hexane and ending with1:1 EtOAc-hexane to furnish5-bromo-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-thiophenesulfonamideas an amorphous white solid (4.3 g, 70.49%). Mass Spectrum (−ESI): 246(M−H)⁻.

B.N-[(1S,2S)-1-(Hydroxymethyl)-2-methylbutyl]-5-(tributylstannyl)-2-thiophenesulfonamide

Bis(Tributyltin) (9.28 mL, 18.52 mmol) and tetrakis(triphenylphosphine)palladium (0) (0.7133 g, 0.617 mmol) were added to a solutionof5-bromo-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-thiophenesulfonamide(4.2 g, 12.34 mmol) in 1,4-dioxane (42 mL). The brown solution washeated to reflux overnight (19 h). An aliquot was taken and TLC (1:1EtOAc-hexane) indicated that reaction was complete. The slurry was thenfiltered and the solvent removed in vacuo to obtain a crude yellow oil(2.1 g). The crude product was purified by column chromatography, silicagel 230-400 mesh, eluent: 1:2 EtOAc-hexane to furnishN-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-5-(tributylstannyl)-2-thiophenesulfonamide as a yellow oil (0.88 g, 12.9%). Mass Spectrum (−ESI): 551(M−H)⁻.

C.N-[(1S,2S)-1-(Hydroxymethyl)-2-methylbutyl]-5-iodo-2-thiophenesulfonamide

To a solution ofN-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-5-(tributylstannyl)-2-thiophenesulfonamide (0.35 g, 0.633 mmol) in methanol (4 mL) was addedsequentially sodium acetate (0.104 g, 1.27 mmol), sodium iodide (0.190g, 1.27 mmol in H₂O) and Chloramine T trihydrate (0.36 g, 1.27 mmol inmethanol (0.5 mL)). The light-yellow solution turned red to orange uponaddition of Chloramine T. The reacted was stirred at 25° C. for 2 h andthen quenched by addition of 1M sodium bisulfite (10 mL). After additionof H₂O (10 mL), the aqueous layer was washed with diethyl ether (3×50mL). The organic layer was dried over MgSO₄, filtered, concentrated toobtain a light yellow oil (0.210 g). The crude product was purified byHPLC (sil(25×0.46 cm); flow rate, 1.0 mL/min; eluent, 6% MTBE in CH₂Cl₂)to furnishN-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-5-iodo-2-thiophenesulfonamideas a white amorphous solid (0.125 g, 51.02%). Mass Spectrum (−ESI): 388(M−H)⁻. Anal. Calc'd for C₁₁H₁₇NIO₄S₂.0.07 EtOAc: C, 31.58; H, 4.21; N,3.64. Found: C, 31.22; H, 4.22; N, 3.54.

Example 2085-Fluoro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-thiophenesulfonamide

A.N-[(1S,2S)-1-(Hydroxymethyl)-2-methylbutyl]-5-(trimethylstannyl)-2-thiophenesulfonamide

Hexamethylditin (5.055 g, 15.43 mmol) and tetrakis(triphenylphosphine)palladium (0) (0.7133 g, 0.617 mmol) were added to a solutionof5-bromo-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-thiophenesulfonamide(prepared as in Example 199, Part A) (3.5 g, 10.27 mmol) in 1,4-dioxane(70 mL). The brown solution was heated to reflux overnight (19 h). Analiquot was taken and TLC (1:1 EtOAc-hexane) indicated that reaction wascomplete. The slurry was then filtered and the solvent removed in vacuoto obtain a crude yellow oil (2.1 g). The crude product was purified bycolumn chromatography, silica gel 230-400 mesh, eluent: 1:2 EtOAc-hexaneto obtainN-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-5-(trimethylstannyl)-2-thiophenesulfon-amideas a yellow oil (3.1 g, 70.8%). Mass Spectrum (−ESI): 425 (M−H)⁻. Anal.Calc'd for C₁₁H₁₇NIO₄S₂: C, 36.64; H, 5.91; N, 3.29. Found: C, 36.64; H,5.81; N, 3.21.

B.5-Fluoro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-thiophenesulfonamide

A solution ofN-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-5-(trimethylstannyl)-2-thiophenesulfonamide (1.0 g, 2.34 mmol) in dry acetonitrile (20 mL) was stirredunder nitrogen at 25° C. Selectfluor (0.850 g, 2.40 mL) was added in oneportion and the solution stirred for 19 h at 25° C. After 3 h a whiteprecipitate began to appear. An aliquot was taken and TLC (1:1EtOAc-hexane) indicated that reaction was not complete. Mainly, startingmaterial was present. The reaction was heated to 80° C. for 6 h. Analiquot was taken and TLC (1:1 EtOAc-hexane) indicated that reaction wascomplete. The slurry was then filtered and the solvent removed in vacuoto obtain a crude yellow oil (0.6 g). The crude product was purified bycolumn chromatography, silica gel 230-400 mesh, eluent: 1:2 EtOAc-hexaneto obtain 5-fluoro-N-[(1S,2S)-1-(hydroxymethyl)-2-methyl-butyl]-2-thiophenesulfonamide as an amorphous white solid (0.102 g,15.49%). Mass Spectrum (−ESI): 280 (M−H)⁻. Anal. Calc'd forC₁₀H₁₆NFO₄S₂: C, 42.69; H, 5.73; N, 4.98. Found: C, 42.47; H, 5.74; N,4.87.

Example 2094-[1-(5-Chloro-thiophene-2-sulfonylamino)-2-hydroxy-ethyl]-piperidine-1-carboxylicacid tert-butyl ester

A. t-Butyl4-((1S)-1-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}-2-hydroxyethyl)piperdine-1-carboxylate

To a solution of cyanuric chloride (1.44 g, 7.80 mmol) in DME (40 mL)was added N-methyl morpholine (0.79 g, 7.80 mmol) at 25° C. A whiteprecipitate formed and to this mixture4-[carboxy-(9H-fluoren-9-ylmethoxycarbonylamino)-methyl]-piperidine-1-carboxylicacid tert-butyl ester (3.75 g, 7.80 mmol) was added as a solutiondissolved in DME (20 mL). After 5 h, the mixture was filtered and theliquid filtrate was cooled in an ice bath to 0° C. and NaBH₄ (0.44 g,11.63 mmol) previously dissolved in H₂O (15 mL) was added by pipet. Thereaction mixture was stirred for an additional 20 min at 0° C. Diethylether (100 mL) was added followed by acidification using 1N HClsolution. The organic phase was then separated and washed with a 10%solution of Na₂CO₃ followed by brine and then dried over MgSO₄.Filtration and evaporation produced a crude glass that was flashchromatographed using ethyl acetate-hexane, 1-1 as eluent. This gave thedesired product as a solid (1.03 g, 28%). MS (+ESI) 367.1 ([M+H]⁺);282.2; 189.1.

B. t-Butyl 4-[(1S)-1-amino-2-hydroxyethyl]-1-piperidinecarboxylate

To tert-butyl4-((1S)-1-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}-2-hydroxyethyl)piperdine-1-carboxylate(0.95 g, 2.03 mmol) was added 20% piperidine in dimethylformamide (20mL) all at once. The reaction was stirred overnight at 25° C. Thedimethylformamide was evaporated off and the crude residue was subjectedto flash chromatography using methylene chloride-methanol-ammoniumhydroxide, 95-5-0.1% as eluent. This yielded the amine product as an oilthat crystallized upon standing (0.392 g, 80%). MS (+ESI) 245.2([M+H]⁺); 189.2; 150.2.

C.4-[1-(5-Chloro-thiophene-2-sulfonylamino)-2-hydroxy-ethyl]-piperidine-1-carboxylicacid tert-butyl ester

To a stirred mixture of tert-butyl4-[(1S)-1-amino-2-hydroxyethyl]-1-piperidinecarboxylate (0.107 g, 0.44mmol), triethylamine (0.046 g, 0.46 mmol) and methylene chloride (5 mL)cooled to 0° C., was added 5-chlorothiophene-2-sulfonyl chloride (0.095g, 0.44 mmol) as a solution dissolved in 2 mL methylene chloride,dropwise by pipet. After 15 min, the ice bath was-removed and thereaction allowed to attain 25° C. and stir overnight. The reaction wasquenched by pouring it into saturated sodium bicarbonate solution (25mL) and additional methylene chloride (15 mL). The organic phase wasseparated and washed sequentially with 1N HCl solution, H₂O, brine anddried over MgSO₄. The organic phase was filtered and evaporated toproduce a crude oil that was flash chromatographed using ethylacetate-hexane, 1-1 as eluent. This produced the title compound as asolid (0.109 g, 58%). MS (+APCI) 442.18 ([M+NH₄]⁺); 386.08; 357.01;325.07; 307.01; 285.06. Anal. Calc'd for C₁₆H₂₅ClN₂O₅S₂: C, 45.22; H,5.93; N, 6.59; Found: C, 45.31; H, 5.87; N, 6.44.

Example 210N-[(1S,2S)-1-(Hydroxymethyl)-2-methylbutyl]thiophene-2-sulfonamide

To a solution of 2-thiophenesulfonyl chloride (1 g, 5.48 mmol) in CH₂Cl₂(5 mL) and (S)-isoleucinol (642 mg, 5.48 mmol) was added Hunig's base(1.05 mL, 6.02 mmol). The reaction mixture was stirred at 25° C. for 24h. The solvent was removed and the oil was dissolved in EtOAc (100 mL).The solution was washed with water (2×100 mL), brine (1×100 mL), anddried over Na₂SO₄. The desired sulfonamide (m/z=264.0(M+H), rt=0.79 min)was isolated by semi-preparative RP-HPLC using the conditions outlinedfor example 195.

Example 2115-Chloro-N—[(S)-2-hydroxy-1-(4-benzylaminocyclohexyl)ethyl]thiophene-2-sulfonamide

A. Part 1

A solution of(S)-2-(5-Cl-thiophenesulfonamido)-2-(4-hydroxycyclohexyl)-N,O-acetonide(4.8 g, 12.7 mmol, see example 199 part 1-3) in CH₂Cl₂ (50 mL) was addedto a slurry of PCC (5.46 g, 25.3 mmol), silica gel (5.46 g) and sodiumacetate (1 g, 12.2 mmol) in CH₂Cl₂ (30 mL). The resulting reactionmixture was stirred at 25° C. overnight. The mixture was diluted withEt₂O and filtered. The solid was washed with diethylether (3×50 mL) andthe combined organic extracts were dried over MgSO₄. The solvent wasremoved in vacuo, and the residue was purified by column chromatographyusing 1:1 EtOAc/Hexane as the eluent to give the ketone as a white solid(4 g, 84%) (100% purity).

B. Part 2

To a solution of(S)-2-(5-Cl-thiophenesulfonamido)-2-(4-cyclohexanone)-N,O-acetonide (340mg, 0.9 mmol) in 1,2-dichloroethane (6 mL) was added benzylamine (118μL, 1.08 mmol), sodium triacetoxyborohydride (286 mg, 1.35 mmol), andacetic acid (52 μL, 0.9 mmol). The reaction was stirred at 25° C.overnight whereupon the reaction was quenched with aqueous NaHCO₃ andextracted with diethyl ether and evaporated. To the resulting residuewas slowly added acetic acid (10 mL of 80%) and the reaction was heatedat 40 ° C. for nine days. The acetic acid was removed and the residuepurified by column chromatography (MeOH/CH₂Cl₂/0.5-1% NH₄OH) to furnishthe desired compound (254 mg, 66%) as a mixture of diastereomers.

The following compounds (Examples 211-220, Table 20) were prepared using(S)-2-(5-Cl-thiophenesulfonamido)-2-(4-cyclohexanone)-N,O-acetonide(from Example 211, part 1) with benzylamine, methylamine, ethylamine,propylamine, allylamine, 3-(aminomethyl)pyridine, morpholine,4-(aminomethyl)pyridine, 2-(aminomethyl)pyridine, and glycine ethylester as outlined in Example 211. TABLE 20 (LCMS Data³: Molecular ionand retention time)

R—NH₂ benzylamine Ex. 211 427.2(M − H), 0.62 min 427.2(M − H), 0.66 minmethylamine Ex. 212 351.2(M − H), 0.39 min 351.1(M − H), 0.45 minethylamine Example 213 365.2(M − H), 0.47 min 365.2(M − H), 0.53 minn-propylamine Ex. 214 379.2(M − H), 0.55 min 379.2(M − H), 0.59 minallylamine Ex. 215 376.9(M − H), 0.42 min 376.9(M − H), 0.47 min3-(aminomethyl) Ex. 216 pyridine 427.9(M − H), 0.35 min 427.9(M − H),0.41 min morpholine Ex. 217 406.9(M − H), 0.40 min 406.9(M − H), 0.44min 4-(aminomethyl) Ex. 218 pyridine 428.0(M − H), 0.49 min 428.0(M −H), 0.51 min 2-(aminomethyl) Ex. 219 pyridine 428.0(M − H), 0.36 min428.0(M − H), 0.37 min glycine ethyl ester Ex. 220 423.0(M − H), 0.51min

Example 221A Method 15-Chloro-N—[(S)-2-ethyl-1-formylbutyl]thiophene-2-sulfonamide

A. 5-(1-Ethyl-propyl)-imidazolidine-2,4-dione

Sodium cyanide (12.0 g, 244.8 mmol) and 2-ethylbutyraldehyde (10.0 mL,81.3 mmol) were added to ammonium carbonate (25.4 g, 325.3 mmol) in H₂O(300 mL). Ethanol (300 mL) was added and salts precipitated. Thereaction mixture was heated to 90° C. After 1 h, the mixture becamehomogeneous and was stirred at 90° C. for 18 h. After cooling to 25° C.,about 500 mL of solvent was removed in vacuo. Concentrated HCl was addedto acidify the mixture to pH 1-2 and a precipitate formed. It wasfiltered and the precipitate was recrystallized from EtOAc to afford5-(1-ethyl-propyl)-imidazolidine-2,4-dione as a white solid (12.9 g,93%). Mass Spectrum (−ESI): 169 (M−H)⁻.

B. N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethylnorvaline

5-(1-Ethyl-propyl)-imidazolidine-2,4-dione (12.3 g, 72.3 mmol) wasdissolved in a 150 mL solution of aqueous NaOH (11.6 g, 289.2 mmol). Thesolution was heated by microwave in a sealed vessel for 1 h. (Microwaveconditions: 15 min @ 100% power, 150° C., 50 psi, then 5 min 0% power,then 15 min @ 100% power, 150° C., 50 psi, then repeat sequence.) Waterand ammonium hydroxide were removed from the reaction mixture in vacuoand the resulting crude amino acid and NaOH mixture was used in the nextreaction without further purification.

The crude amino acid and NaOH mixture was dissolved in 300 mL of water.The mixture was cooled to 0° C. in an ice bath.5-Chlorothiophene-2-sulfonyl chloride (17.3 g, 79.5 mmol) was dissolvedin 100 mL of THF and added dropwise to the reaction mixture over 0.5 h.After 1 h the reaction mixture was allowed to warm gradually to 25° C.and stirred for 16 h. THF was removed in vacuo and then the mixture wasacidified to pH 1 with 1N HCl. After about 15 min, a precipitate beganto crash out of the milky white solution. After 1 h, the mixture wascooled in a refrigerator for 1 h and then filtered. The precipitate waswashed with 1 N HCl to provideN-[(5-chloro-2-thienyl)sulfonyl]-3-ethylnorvaline as a white solid (18.5g, 78%). Mass Spectrum (−ESI): 325 (M−H)⁻.

C. N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline

(+)-(1S,2R)-Ephedrine hemihydrate (16.7 g, 95.6 mmol) was added to asuspension of N-[(5-chloro-2-thienyl)sulfonyl]-3-ethylnorvaline (31.2 g,95.6 mmol) in 185 mL of EtOH. The mixture was heated slightly todissolve solids and a precipitate formed. After cooling at 5° C. for 18h the resulting-suspension was filtered and the precipitate was washedwith cold EtOH and EtOAc to give 27% yield of the diastereomeric salt.The salt was recrystallized from boiling EtOAc (420 mL), then filteredoff. The resulting white solid was then dissolved in 300 mL of EtOAc and300 mL of 1N HCl. The layers were separated and the organic extract waswashed with 1N HCl (2×200 mL), dried (Na₂SO₄), and concentrated to giveN-[(5-chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline as a white solid(5.6 g, 18%). Chiral HPLC [chiralpak AD (25×0.46 cm), 8:2 hexane (0.1%TFA) : isopropanol, L-isomer elutes at 9.6 min and D-isomer elutes at13.1 min] indicated 96% chiral purity. [α]_(D) ²⁵=+44.5° (c=1% SOLUTION,MeOH). Mass Spectrum (−ESI): 325 (M−H)⁻. Anal. Calc'd for C₁₁H₁₆ClNO₄S₂:C, 40.55; H, 4.95; N, 4.30. Found: C, 40.30; H, 4.78; N, 4.16.

D.5-Chloro-N—[(S)-2-ethyl-1-(hydroxymethyl)butyl]-2-thiophenesulfonamide

To N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline (5.6 g, 17.2mmol) in THF (150 mL) at 0° C. was added a solution of 1 M boranetetrahydrofuran complex in THF (69 mL, 69 mmol) dropwise via additionfunnel. After 15 min, the reaction mixture was warmed to 25° C. andstirred for 18 h. It was then quenched with 90 mL of 10% AcOH in MeOHslowly. Volatiles were removed in vacuo. The residue was then dissolvedin EtOAc (300 mL) and washed with sat. aqueous NaHCO₃ (3×200 mL), dried(Na₂SO4), and concentrated to a white precipitate (5.1 g, 96% yield, 96%chiral purity). The precipitate was recrystallized with heptane/EtOAc,4:1, to give optically pure5-chloro-N—[(S)-2-ethyl-1-(hydroxymethyl)butyl]-2-thiophenesulfonamideas white needles (4.4 g, 81% yield). [α]_(D) ²⁵=+4.5° (c=1% SOLUTION,DMSO). Mass Spectrum (−ESI): 310 (M−H)⁻. Anal. Calc'd for C₁₁H₁₈ClNO₃S₂:C, 42.37; H, 5.82; N, 4.49. Found: C,42.37; H, 5.79; N, 4.38.

E. 5-Chloro-N—[(S)-2-ethyl-1-formylbutyl]thiophene-2-sulfonamide

Pyridinium dichromate (2.4 g, 6.4 mmol) was added to a solution of5-chloro-N—[(S)-2-ethyl-1-(hydroxymethyl)butyl]-2-thiophenesulfonamide(0.5 g, 1.6 mmol) in CH₂Cl₂ (20 mL). After 18 h, the reaction mixturewas filtered through a plug of Celite. The filtrate was concentrated andthe resulting residue was purified by silica gel column chromatography(eluant: 1:4 EtOAc-hexane) to give5-chloro-N—[(S)-2-ethyl-1-formylbutyl]thiophene-2-sulfonamide as a whitesolid (303 mg, 61%). [α]_(D) ²⁵=+136.76° (c=1% SOLUTION, CHCl₃). MassSpectrum (−ESI): 308 (M−H)⁻. Anal. Calc'd for C₁₁H₁₆ClNO₃S₂: C, 42.64;H, 5.21; N, 4.52. Found: C, 42.57; H, 5.24; N, 4.52.

Example 221B Method 25-Chloro-N—[(S)-2-ethyl-1-formylbutyl]thiophene-2-sulfonamide A.(S)-3-Ethyl-2-{[(S)-1-phenylethyl]amino}pentanenitrile

To (S)-(−)-α-methylbenzylamine hydrochloride salt (1.2 g, 7.6 mmol) in80 mL of 1:1 MeOH/H₂O was added potassium cyanide (0.5 g, 7.6 mmol) and2-methylbutyraldehyde (0.94 mL, 7.6 mmol). A precipitate formed after 30min. After 20 h, the suspension was filtered and washed with H₂O to give(S)-3-ethyl-2-{[(S)-1-phenylethyl]amino}pentanenitrile as a white powder(1.29 g, 74%). Mass Spectrum (+ESI): 310 (M+H)⁺. Anal. Calc'd forC₁₅H₂₂N₂: C, 78.21; H19.63; N, 12.16. Found: C, 77.90; H, 9.75; N,12.32.

B. 3-Ethyl-N²-[(S)-1-phenylethyl]-L-norvalinamide

To 25 mL of sulfuric acid at 0° C. was added(S)-3-ethyl-2-{[(S)-1-phenylethyl]amino}pentanenitrile (2.7 g, 11.6mmol) portionwise. The mixture was warmed to 25° C. After 2 days, thereaction mixture was poured over about 100 g of crushed ice.Concentrated NH₄OH was added to neutralize the acid. This mixture wasextracted with EtOAc (3×100 mL), dried over Na₂SO₄, filtered andconcentrated to give 3-ethyl-N²-[(S)-1-phenylethyl]-L-norvalinamide (2.6g, 90%), which was used in the next step without purification. MassSpectrum (+ESI): 249 (M+H)⁺. Anal. Calc'd for C₁₅H₂₄N₂O: C, 72.54; H,9.74; N, 11.28. Found: C, 72.24; H, 10.04; N, 11.01

C. 3-Ethyl-L-norvalinamide

A mixture of 3-ethyl-N²-[(S)-1-phenylethyl]-L-norvalinamide (2.6 g, 10.5mmol) and 5% Pd/C (800 mg) was shaken for 24 h in a Parr apparatus under3 atm of H₂. The mixture was filtered through a plug of Celite and thesolvent was removed in vacuo to give 3-ethyl-L-norvalinamide as a whitesolid (1.4 g, 93%), which was used in the next reaction without furtherpurification. Mass Spectrum (+ESI): 145 (M+H)⁺.

D. N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline

3-Ethyl-L-norvalinamide (1.2 g, 4.8 mmol) was dissolved in conc. HCl (10mL) and heated to 100° C. for 16 h. The reaction mixture wasconcentrated to a white solid consisting of the amino acid hydrochloridesalt and one equivalent of NH₄Cl, which was used in the next reactionwithout purification.

Amino acid hydrochloride salt with 1 equivalent of NH₄Cl (0.28 g, 1.19mmol) was dissolved in 6 mL of H₂O and then NaOH (0.24 g, 6.00 mmol) wasadded. The solution was cooled to 0° C. and then5-chlorothiophene-2-sulfonyl chloride (0.29 g, 1.32 mmol) in 6 mL of THFwas added dropwise. The mixture was warmed to 25° C. After 19 h, THF wasremoved in vacuo. The remaining solution was diluted with 10 mL of H₂Oand washed with EtOAc (2×10 mL). The solution was acidified with 1N HCland a precipitate formed. This was filtered to giveN-[(5-chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline as a white solid(0.17 g, 44%). Chiral HPLC indicates that only the S enantiomer ispresent.

5-Chloro-N—[(S)-2-ethyl-1-(hydroxymethyl)butyl]-2-thiophenesulfonamideand 5-Chloro-N—[(S)-2-ethyl-1-formylbutyl]thiophene-2-sulfonamide werethen prepared from N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvalineaccording to method 1 of Example 221A.

Example 221C5-Chloro-N—[(S)-2-ethyl-1-(hydroxymethyl)butyl]-2-thiophenesulfonamide

Into a 3L 3-necked flask equipped with a nitrogen inlet tube, amechanical stirrer, and an addition funnel with a stopper was placedlithium borohydride (145 mL of a 2 M solution in THF, 0.29 mol). Thesolution was placed under nitrogen and cooled to 0° C.Chlorotrimethylsilane (73.8 mL, 0.58 mol) was added dropwise over aperiod of 30 min. The ice bath was removed and the resulting slurry wasstirred at room temperature for 30 min. The reaction mixture was cooledto 0° C. and the 2-(S)-amino-3-ethyl-pentanoic acid (21.1 g, 0.145 mol),which was prepared according to Scheme 13, was added in portions as asolid over a period of 15 min. The reaction mixture was allowed to warmslowly to room temperature as the ice bath melted. After 3 days at roomtemperature, the reaction mixture was cooled to 0° C., and methanol (217mL) was carefully added over a period of 80 min. The solution wasstirred at room temperature for an additional 40 min, then concentratedunder reduced pressure in a water bath at 60° C. The resulting slurrywas made basic with 20% sodium hydroxide (37.5 mL). Water (37.5 mL) wasadded, and the entire aqueous layer was extracted with methylenechloride (300 mL), and dried (Na₂SO₄). Concentration under reducedpressure gave 2(S)-amino-3-ethylpentanol as an oil (17.3 g, 91%), whichwas used immediately or stored in the freezer overnight: Opt. Rot.[α]_(D) ²⁵=−3.7° (1% solution, DMSO); ¹H NMR (DMSO-d⁶, 500 MHz): δ 4.38(broad s, 1H), 3.35 (dd overlapping with a broad s at δ 3.32, J=4.5,10.3 Hz, 3H), 3.14 (dd, J=7.9, 10.2 Hz, 1H), 2.63 (m, 1H), 1.45-1.05 (m,5H), 0.82 and 0.81(two overlapping triplets, J=7.4 Hz, 6H); MS(+ESI):[M+H]⁺, 132 (60%).

A mixture of 2(S)-amino-3-ethylpentanol (34.1 g, 0.26 mol) and methylenechloride (700 mL) was placed under Argon, and cooled to 0° C.Triethylamine (36.2 mL, 0.26 mol) was added, followed by the dropwiseaddition of 5-chlorothiophene-2-sulfonyl chloride (56.4 g, 0.26 mol) inmethylene chloride (400 mL). The reaction mixture was allowed to warmslowly to room temperature as the ice bath melted. After 3 days at roomtemperature, the reaction mixture was divided into two-0.6 L portions.Each portion was diluted with ethyl acetate (1L), and washed three timeswith saturated potassium phosphate monohydrate (200 mL), once with brine(200 mL), and dried (Na₂SO₄). Concentration under reduced pressure gavea white solid (74.5 g, 92%). The product (87.98 g) from several runswere combined and recrystallized from hot heptane:ethyl acetate (4:1,775 mL) to give the title compound as crystals (74.9 g, 85%): mp115-117.6° C.; Opt. Rot. [α]_(D) ²⁵=+10.81° (1% solution, MeOH); ¹H NMR(DMSO-d⁶, 500 MHz): δ 7.71 (d, J=8.1 Hz, 1H), 7.44 (d, J=4.1 Hz, 1H),7.22 (d, J=4.1 Hz, 1H), 4.56 (t, J=5.2 Hz, OH), 3.31-3.15 (m, 3H),1.40-1.115 (m, 4H), 1.07 (m, 1H), 0.79 and 0.76 (two overlappingtriplets, J=7.3 Hz, 6H); ¹³C NMR (DMSO-d⁶, 100 MHz): δ 141.75, 133.73,130.95, 127.60, 60.41, 56.89, 41.57, 21.31, 20.80, 11.79, 11.51;MS(−ESI): [M−H]⁻, 1 chlorine isotope pattern, 310 (100%), 312 (30%);Anal. Calc. for C₁₁H₁₈ClNO₃S₂: C, 42.37, H, 5.82, N, 4.49. Found: C,42.34, H, 5.65, N, 4.43. Chiral HPLC (Chiralpak A D, 25×0.46 cm, eluant8:2 hexane/isopropanol containing 0.1% TFA, flow rate 0.5 mL/min, UVdetection at 254 nm, retention times for the S and R isomers are 10.95min and 11.95 min, respectively) revealed an S/R ratio of 100.0:0.0.

Example 2225-Chloro-N—[(S)-2-ethyl-1-(1-hydroxyethyl)butyl]thiophene-2-sulfonamide

A solution of methylmagnesium bromide (1.4 M, 7.0 mL, 9.7 mmol) intoluene/THF (75:25) was added to a 0° C. solution of5-chloro-N—[(S)-2-ethyl-1-formylbutyl]thiophene-2-sulfonamide (Example221, 1.0 g, 3.2 mmol) in THF (30 mL). The mixture was warmed to 25° C.and after 2 h was quenched carefullly with saturated aqueous ammoniumchloride (25 mL). The mixture was extracted with EtOAc (3×25 mL). Theorganic extract was dried over Na₂SO₄, filtered and concentrated to givea colorless oil. The product was purified by column chromatography(Biotage), eluant: 1:4 EtOAc-hexane, to afford5-chloro-N—[(S)-2-ethyl-1-(1-hydroxyethyl)butyl]thiophene-2-sulfonamideas a white solid (876 mg, 83%). The product is a diastereomeric mixturewith a ratio of 3:7. mp 95-98° C. Anal. Calc'd for C₁₂H₂₀ClNO₃S₂: C,44.23; H, 6.19; N, 4.30. Found: C, 44.25 H, 6.35; N, 4.29. Mass Spectrum(−ESI): 324 (M−H)⁻.

Example 2235-Chloro-N—[(S)-2-ethyl-1-(1-hydroxy-1-methylethyl)butyl]thiophene-2-sulfonamide

A. N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline methyl ester

Trimethylsilyldiazomethane (3.1 mL, 6.1 mmol) was added to a solution ofN-[(5-chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline (1.0 g, 3.1 mmol)in THF (20 mL) and MeOH (5 mL). After 2 h the mixture was concentratedto give N-[(5-chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline methylester as a white solid (1.0 g, 99%). Mass Spectrum (−ESI): 338.00(M−H)⁻.

B.5-Chloro-N—[(S)-2-ethyl-1-(1-hydroxy-1-methylethyl)butyl]thiophene-2-sulfonamide

A solution of methylmagnesium bromide (1.4 M, 9.5 mL, 13.2 mmol) intoluene/THF (75:25) was added to a 0° C. solution ofN-[(5-chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline methyl ester (0.90g, 2.65 mmol) in THF (26 mL). The solution was allowed to warm to 25°C., then heated to 55° C. and stirred for 18 h. It was then cooled to 0°C. and quenched slowly with saturated aqueous NH₄Cl. EtOAc (75 mL) wasadded and the phases were separated. The organic layer was dried overNa₂SO₄, filtered and concentrated to give a yellow oil. The crudeproduct was purified by column chromatography (Biotage), eluant: 1:4EtOAc-hexane, to afford5-chloro-N—[(S)-2-ethyl-1-(1-hydroxy-1-methylethyl)butyl]thiophene-2-sulfonamideas a colorless oil (0.72 g, 80%). Mass Spectrum (−ESI): 338 (M−H)⁻.Anal. Calc'd for C₁₃H₂₂ClNO₃S₂: C, 45.94; H, 6.52; N, 4.12. Found: C,46.10; H, 6.63; N, 4.04.

Example 2245-Chloro-N-(2-hydroxy-1-tetrahydro-H-thiopyran-4-ylethyl)thiophene-2-sulfonamide

A.(5-Chloro-thiophene-2-sulfonylamino)-(tetrahydro-thiopyran-4-yl)-aceticacid

Sodium hydroxide (0.20 g, 5.04 mmol) was added to a mixture ofN-Fmoc-amino-(4-tetrahydrothiopyranyl)acetic acid (0.50 g, 1.26 mmol) inMeOH:water, 2:1 (15 mL) at 25° C. The reaction mixture was allowed tostir for 20 h. TLC (1:9 MeOH/CHCl₃) indicated that the reaction wascomplete. The mixture was diluted with water and washed with EtOAc. Thewater layer was concentrated to give a white solid with NaOH remaining.This white solid was redissolved in H₂O:THF, 1:2 (15 mL) and cooled to0° C. 5-chlorothiophene-2-sulfonyl chloride (0.683 g, 3.15 mmol) wasdissolved in THF (2 mL) and added to the mixture dropwise which was thenwarmed to 25° C. overnight. Aqueous 1 N HCl was added to acidify themixture to pH 1. EtOAc was added and the layers were separated. Theorganic extract was washed with 1 N HCl, and H₂O, dried over Na₂SO₄,filtered and concentrated to give(5-chloro-thiophene-2-sulfonylamino)-(tetrahydro-thiopyran-4-yl)-aceticacid as a reddish black solid (0.14, 31%), which was used in the nextreaction without purification. Mass Spectrum (+ESI): 357 (M+H)⁺.

B.5-Chloro-N-(2-hydroxy-1-tetrahydro-H-thiopyran-4-ylethyl)thiophene-2-sulfonamide

(5-Chloro-thiophene-2-sulfonylamino)-(tetrahydro-thiopyran-4-yl)-aceticacid (0.14 g, 0.40 mmol) was dissolved in THF (2 mL) and cooled to 0° C.A solution of borane tetrahydrofuran complex (1 M, 3.2 mL, 3.2 mmol) inTHF was added dropwise and the mixture was allowed to warm to 25° C.overnight. The volatile solvents were removed in vacuo and the resultingorange oil was diluted with EtOAc and washed with H₂O, 1 N HCl, andsaturated aqueous NaHCO₃. The organic extract was dried over Na₂SO₄,filtered and concentrated. The resulting residue was purified by columnchromatography (Biotage), eluant: 1:1 EtOAc:hexane, to afford5-chloro-N-(2-hydroxy-1-tetrahydro-H-thiopyran-4-ylethyl)thiophene-2-sulfonamide(40 mg, 30%) as a white solid. mp 108-110° C. Mass Spectrum (−ESI): 340(M−H)⁻. Anal. Calc'd for C₁₁H₁₆ClNO₃S₃: C, 38.64; H, 4.72; N, 4.10.Found: C, 38.80; H, 4.69; N, 3.88.

Example 2255-Chloro-N—[(S)-2-hydroxy-1-piperidin-4-ylethyl]thiophene-2-sulfonamide

To a solution of4-[1-(5-chloro-thiophene-2-sulfonylamino)-2-hydroxy-ethyl]-piperidine-1-carboxylicacid tert-butyl ester (0.204 g, 0.48 mmol (see example 209)) dissolvedin dichloromethane (2 mL) at 0° C. was added trifluoroacetic acid (0.5mL). The reaction was allowed to warm to 25° C. and stir overnight. Themixture was then concentrated and dichloromethane was added andevaporated 6 times to yield a crude solid. Purification by HPLC (C-18column, (21×75 mm) with elution system 60-100% acetonitrile-water+0.1%TFA, 20 min gradient) gave the product as an oil (0.0166 g, 11%). MS(ESI) m/z 325 ([M+H]⁺).

Example 226N—[(S)-2-Ethyl-1-(hydroxymethyl)butyl]thiophene-2-sulfonamide

A.N-[(1S)-2-Ethyl-1-(hydroxymethyl)butyl]-5-(trimethylstannyl)thiophene-2-sulfonamide

A solution of5-bromo-N-[(1S)-2-ethyl-1-(hydroxymethyl)butyl]thiophene-2-sulfonamide(0.71 g, 2.0 mmol), hexamethylditin (0.983 g, 3.0 mmol),tetrakis(triphenylphosphine)palladium (0.115 g, 0.10 mmol) and1,4-dioxane (15 mL) was refluxed for 16 h under a nitrogen atmosphere.After cooling to 25° C., dichloromethane (10 mL) was added and themixture filtered and evaporated to give the product as a crude oil (0.49g), that was used without purification in the next step, part B. MS(−ESI) 439.20 ([M−H]−).

B. N—[(S)-2-Ethyl-1-(hydroxymethyl)butyl]thiophene-2-sulfonamide

To a stirred mixture of anhydrous acetonitrile (6 mL) andN-[(1S)-2-ethyl-1-(hydroxymethyl)butyl]-5-(trimethylstannyl)thiophene-2-sulfonamide(0.24 g, 0.56 mmol) was added Selectfluor (Aldrich) (0.204 g, 0.57 mmol)all at once. The mixture was heated to 75° C. under a nitrogenatmosphere, stirred 16 h and then cooled to 25° C. and filtered.Evaporation of the solvent produced a crude solid that was taken up inethyl acetate and again filtered to remove insoluble solids. Evaporationof the remaining solvent produced an oil that was purified by flashchromatography using hexane-ethyl acetate 2-1 as eluant, producing thetitle compound as the major product (0.051 g, 33%). MS (−ESI) 276.20([M−H]⁻).

Example 227N—[(S)-2-Ethyl-1-(hydroxymethyl)butyl]-5-fluorothiophene-2-sulfonamide

This compound was synthesized as a side product using the procedurefound in example 226 (parts A and B) and was isolated from the sameflash chromatography column as a solid (0.024 g, 15%). MS (−ESI) 294.20([M−H]⁻).

Example 2285-Chloro-N-[(1S)-1-(2,3-dihydro-1H-inden-2-yl)-2-hydroxyethyl]thiophene-2-sulfonamide

A.9H-Fluoren-9-ylmethyl-(1S)-1-(2,3-dihydro-1H-inden-2-yl)-2-hydroxyethylcarbamate

1N Borane-THF (24.18 mL) was added dropwise over 30 min at 0° C. to asolution of(2S-2,3-dihydro-1H-indene-2-yl[[(9H-fluoren-9-ylmethoxy)carbonyl]amino)ethanoicacid (2.0 g, 4.84 mmol) in anhydrous tetrahydrofuran (20 mL). Thereaction was allowed to warm to 25° C. overnight, and then was quenchedby addition of 10.0 mL of 10% acetic acid in methanol. After solventevaporation, the crude product was dissolved in ethyl acetate and washedwith 1N HCl, water and 10% NaHCO₃. The organic layer was dried overMgSO₄, filtered and concentrated to obtain a crude yellow oil (1.8 g).The crude product was purified by column chromatography, silica gel230-400 mesh, eluant: 1:2 EtOAc-hexane. This produced the title compoundas an amorphous solid (1.05 g, 54.4%). Mass Spectrum (−ESI): 398 (M−H)⁻.(+ESI): 400 (M+H)⁺.

B. (2S)-2-Amino-2-(2,3-dihydro-1H-inden-2-yl)ethanol

20% Piperidine in DMF (15 mL) was added to a solution of9H-fluoren-9-ylmethyl-(1S)-1-(2,3-dihydro-1H-inden-2-yl)-2-hydroxyethylcarbamate(1.05 g, 2.63 mmol) in DMF (5 mL). The reaction was stirred at 25° C.for 19 h. After solvent evaporation, the crude product was dissolved inethyl acetate (50 mL) and dried over MgSO₄, filtered and concentrated toobtain a crude yellow oil (1.05 g). Mass Spectrum (+ESI): 179 (M+H)⁺.

C.5-Chloro-N-[(1S)-1-(2,3-dihydro-1H-inden-2-yl)-2-hydroxyethyl]thiophene-2-sulfonamide

5-Chlorothiophene-2-sulfonyl chloride (0.856 g, 3.94 mmol) was addeddropwise (5 min) as a solution in CH₂Cl₂ (5 mL) to a 0° C. solution of(2S)-2-amino-2-(2,3-dihydro-1H-inden-2-yl)ethanol (0.46 g, 2.63 mmol) inCH₂Cl₂ (5 mL) and triethylamine (3.8 mL, 5.26 mmol). The solution wasallowed to warm to 25° C. overnight (19 h). An aliquot was taken and TLC(1:1 EtOAc-hexane) indicated that reaction was complete. It was dilutedwith CH₂Cl₂ (50 mL) and the organic layer was washed with 1N HCl (2×50mL), saturated aqueous NaCl (50 mL). The organic layer was dried overMgSO₄, filtered and concentrated to obtain a crude oil (0.89 g). Thecrude product was purified by column chromatography, silica gel 230-400mesh, eluant: 1:4 EtOAc-hexane to afford5-chloro-N-[(1S)-1-(2,3-dihydro-1H-inden-2-yl)-2-hydroxy-ethyl]thiophene-2-sulfonamideas an amorphous white solid (0.361 g, 38.4%). Mass Spectrum (−ESI): 356(M−H)⁻. Anal. Calc'd for C₁₅H₁₀ClNO₃S₂: C: 50.34 H: 4.51 N: 3.91 Found:C: 50.28 H: 4.36 N: 3.77.

Example 2295-Chloro-N-{(1S,2S)-1-[(Z)-(hydroxyimino)methyl]-2-methylbutylthiophene-2-sulfonamide

A solution of5-chloro-N-[(1S,2S)-1-formyl-2-methylbutyl]thiophene-2-sulfonamide(Example 118, 1.0 g, 3.4 mmol), hydroxylamine hydrochloride (0.464 g,6.78 mmol) and sodium acetate (0.556 g, 6.78 mmol) in methanol (10 mL)was stirred under reflux for 19 h. After evaporation of the solvent, theresidue was diluted with aqueous K₂CO₃ (20 mL) and then extracted withCH₂Cl₂ (2×40 mL). The combined reaction extracts were washed with brine,dried over MgSO₄, filtered and concentrated to obtain a crude oil (0.89g). The crude product was purified by column chromatography, silica gel230-400 mesh, eluant: 1:4 EtOAc-hexane. This produced the title compound(Z-isomer) as an amorphous white solid (32 mg, 3.1%). Mass Spectrum(−ESI): 309 (M−H)⁻. Anal. Calc'd for C₁₀H₁₅ClN₂O₃S₂. 0.10 C₄H₈O₂: C:39.08 H: 4.98 N: 8.76 Found: C: 38.72 H: 4.67 N: 8.43.

Example 2305-chloro-N—{(S,S)-1-[(E)-(hydroxyimino)methyl]-2-methylbutylthiophene-2-sulfonamide

A solution of5-chloro-N-[(1R,2S)-1-formyl-2-methylbutyl]thiophene-2-sulfonamide(Example 118, 1.0 g, 3.4 mmol), hydroxylamine hydrochloride (0.464 g,6.78 mmol) and sodium acetate (0.556 g, 6.78 mmol) in methanol (10 mL)was stirred under reflux for 19 h. After evaporation of the solvent, theresidue was diluted with aqueous K₂CO₃ (20 mL) and then extracted withCH₂Cl₂ (2×40 mL). The combined reaction extracts were washed with brine,dried over MgSO₄, filtered, and concentrated to obtain a crude oil (0.89g). The crude product was purified by column chromatography, silica gel230-400 mesh, eluant: 1:4 EtOAc-hexane This produced the title compound(E-isomer) as an amorphous white solid (300 mg, 28.3%). Mass Spectrum(−ESI): 309 (M−H)⁻. Anal. Calc'd for C₁₀H₁₅ClN₂O₃S₂.0.40 C₄H₈O₂: C:40.26 H: 5.30 N: 8.09 Found: 39.78 H: 5.23 N: 7.77

A. Diethyl-3-ethyl-5-oxopyrrolidine-2,2-dicarboxylate

To 150 mg of sodium dissolved in 150 mL of absolute ethanol was addeddiethyl acetamidomalonate (5.3 g, 25 mmol) and ethyl-(2E)-pent-2-enoate(3.5 g, 27.3 mmol). The reaction mixture was then refluxed for 20 h.After this period of time, 2 mL of glacial acetic acid was added,volatiles were removed under pressure with the aid of a water aspiratorand heating bath. On cooling the residue solidified. The residue wasdissolved in 50 mL of toluene and to this was added 20 mL of petroleumether. The product precipitated when the mixture was cooled. Thecrystals were collected and washed with water and further dried in vacuoto obtain a white solid (5.6 g, 79.77%). Mass Spectrum (+ESI): 258(M+H)⁺.

B. 3-Ethylglutamic acid

5.6 g of Diethyl-3-ethyl-5-oxopyrrolidine-2,2-dicarboxylate (21.76,84.67 mmol) was refluxed in 80 mL of 49% fuming HBr for 4 h. After thistime, the contents were placed in vacuo and the volatile constituentswere removed. The gummy residue was dissolved in 25 mL of distilledwater and the water was removed as before. The process was repeated oncemore. The residue was dissolved in 20 mL of water and the pH of thesolution was adjusted to pH 3 with concentrated ammonia (2 mL) solution.At this point precipitation of the ethyl glutamic acid was encouraged bycooling on an ice bath or by diluting the aqueous solution with 100 mLof absolute ethanol. Precipitation from the water-ethanol mixture iscomplete in 48 h. Care must be taken to add the ethanol slowly toprevent the precipitation of an undesired side product. The compound waspurified by crystallization from water-ethanol (1:1) mixture. Thisproduced the title compound as an amorphous white solid (3.5 g, 99%).Mass Spectrum (+ESI): 176 (M+H)⁺.

C. 3-Ethyl-2-methylpentane-1,5-diol

To a slurry of LAH (2.06 g, 54.29 mmol) in THF (60 mL) was added3-ethylglutamic acid (3.5 g, 21.71 mmol) dropwise at 0° C. over 20 min.The reaction was heated to 36° C. for 18 h. The reaction slurry (gray)was cooled to 0° C. and quenched with H₂O (3 mL) then washed with 1NNaOH (9 mL) and H₂O (3 mL). It was then stirred for 6 h at 25° C. toobtain an off-white slurry. The slurry was filtered and the motherliquor was further dried over MgSO₄, filtered and concentrated in vacuoto obtain 3-ethyl-2-methylpentane-1,5-diol as a crude yellow oil (2.85g, 89.17%). Mass Spectrum (+ESI): 170 (M+Na)⁺.

D.5-Chloro-N-[2-ethyl-4-hydroxy-1-(hydroxymethyl)butyl]thiophene-2-sulfonamide

5-Chlorothiophene-2-sulfonyl chloride (6.4 g, 24.48 mmol) was addeddropwise (5 min) as a solution in CH₂Cl₂ (5 mL) to a 0° C. solution of3-ethyl-2-methylpentane-1,5-diol (2.85 g, 19.34 mmol) in CH₂Cl₂ (30 mL)and triethylamine (5.66 mL, 40.81 mmol). The solution was allowed towarn to 25° C. overnight (19 h). An aliquot was taken and TLC (1:1EtOAc-hexane) indicated that reaction was complete. It was diluted withCH₂Cl₂ (50 mL) and the organic layer was washed with 1N HCl (2×50 mL)and saturated aqueous NaCl (50 mL). The organic layer was dried overMgSO₄, filtered and concentrated to obtain a crude oil (4.9 g). Thecrude product was purified by column chromatography, silica gel 230-400mesh, eluant: 1:4 EtOAc-hexane to afford5-chloro-N-[2-ethyl-4-hydroxy-1-(hydroxymethyl)butyl]thiophene-2-sulfonamideas an amorphous white solid (0.450 g, 7.3%). Mass Spectrum (−ESI): 326(M−H)⁻.

1. Semi-Preparative RP-HPLC Conditions:

-   Gilson Semi-Preparative HPLC system with Unipoint Software.-   Column: Phenomenex C18 Luna 21.6 mm×60 mm, 5μ-   Solvent A: Water (0.02% TFA buffer)-   Solvent B: Acetonitrile (0.02% TFA buffer)-   Solvent Gradient: Time 0: 10% B; 2.5 min: 10% B; 14 min: 90% B.-   Flow Rate: 22.5 mL/min-   The product peak was collected based on UV absorption and    concentrated.    2. Analytical LCMS Conditions:-   Hewlett Packard 1100 MSD with ChemStation Software-   Column: YMC ODS-AM 2.0 mm×50 mm 5μ column at 23° C.,-   3 μL injection;-   Solvent A: Water (0.02% TFA buffer)-   Solvent B: Acetonitrile (0.02% TFA buffer)-   Gradient: Time 0: 95% A; 0.3 min: 95% A; 4.7 min: 10% A; 4.9 min:    95% A.-   Flow rate 1.5 mL/min;-   Detection: 254 nm DAD;-   API-ES Scanning Mode Positive 150-700; Fragmentor 70 mV.    3. Analytical LCMS Conditions:-   ZMD (Waters) or Platform (Micromass) or LCZ (Micromass)-   Column: Zorbax SB-C8-   Solvent: Acetonitrile+H₂O containing 0.1% TTA or 0.1% FA-   Gradient: Gradient: 2.5 min 15% Acetonitrile—95% Acetonitrile-   Flow rate 3 ml\min-   Detection: ELSD detection (SEDEX 55)-   UV 253 detection (Schimadzu)

Example 233 Repressor Release Assay (RRA)

The compounds generated as described in Examples 1 through 220 weretested in the RRA in accordance with published techniques [Shuey, D. J.,Sheiffele, P., Jones, D., Cockett, M. I., and Quinet, E. M. (1999),“Repressor release: a useful tool for monitoring amyloid precursorprotein (APP) proteolysis in mammalian cells”, Society for NeuroscienceAbstracts, Vol. 25, 29^(th) Annual Meeting of Society for Neuroscience,Miami Beach, Fla., Oct. 23-28, 1999]. Briefly, this assay is performedas follows.

A. Cell Culture

CHO-K1 cells are cultured in whole DMEM media (DMEM—High Glucose with10% fetal bovine serum, 1% Non-essential Amino Acids, and 1%Penicillin-Streptomycin) at 37° C. with 5% CO₂. Two million cells areplated into 10-cm dishes 24 hrs prior to transfection.

Transient transfections are completed as recommended by Gibco BRL usingtheir Lipofectamine Plus system. First, 6 μg of pRSVO-luc and 6 μg ofAPP-lacI construct DNA are added to 460 μL Opti-Mem transfection mediaand incubated with 30 μL Plus reagent for 15 minutes. Then, a lipidmixture of 40 μL Lipofectamine reagent and 460 μL Opti-Mem transfectionmedia is incubated with the DNA-Plus reagent mixture for 15 minutes.During the DNA-lipid incubation, the CHO-K1 cells are washed once andcovered in 5.0 mL DMEM media without Penicillin-Streptomycin. TheDNA-lipid preparation is then layered onto these cells and incubated at37° C. overnight.

One and one half million transfected cells per well (100 μL totalvolume) are plated into sterile, opaque Packard 96-well Cultur-Plates inclear DMEM whole media (DMEM—without phenol red) and incubated at 37° C.with 5% CO₂ for 3-5 hours.

B. Compound Dilution

Compounds are diluted using two different protocols; one protocol isused for compounds supplied neat (weighed powder in vial) and the otherprotocol is used for compounds supplied in solution (20 mM in DMSO in96-well plates). For both protocols, 25 mM Hepes and 25 mM Hepes/1% DMSOare prepared fresh to be used as diluent. The Hepes/DMSO is used as thediluent control on all experimental plates.

The following table depicts the steps for compound dilution (please notethat the last step is the addition of compound to cells/media in tissueculture plate): TABLE 21 Concentration Dilution Stock Solution 10 mg/mLx mg compound (vial) diluted with 100% DMSO Dilution 1 1 mg/mL 20 μLstock solution 180 μL 25 mM Hepes Dilution 2 200 μg/mL 60 μL Dilution 1240 μL 25 mM Hepes Dilution 3 20 μg/mL 11.3 μL Dilution 2 (in CellPlate) (in 100 μL cells/well)

Because some compounds arrive in 96-well format at 20 mM, the followingrepresents the protocol for their dilution (note that an averagemolecular weight of these compounds was used to calculate thesedilutions and as above, the last step is the addition of compound tocells/media in tissue culture plate): TABLE 22 Concentration DilutionStock Solution (original conc.) — 20 mM Solution Dilution 1 ˜200 μg/mL 6μL stock solution 194 μL 25 mM Hepes Dilution 2 (in Cell Plate)  ˜20μg/mL 11.3 μL Dilution 2 (in 100 μL cells/well)

Once compounds are diluted, they are applied in duplicate on cells intissue culture plates (prepared above). Cells are incubated withcompound at 37° C. with 5% CO₂ for an additional 36-48 hours.

C. Assay Measurement

Luciferase assays (LucLite reagent, Packard) are performed and are readon a Packard TopCount instrument. Media is removed from each 96-wellplate and replaced with 100 μL PBS per well (with Mg²⁺ and Ca²⁺). Anequal volume (100 μL) of the LucLite lysis/substrate buffer is added toeach well and the plates are sealed and mixed in the dark on a rotaryshaker for 15-30 minutes at room temperature. Luciferase readings arethen taken on the TopCount instrument. Measurements are expressed asrelative light units (RLU) and are calculated and analyzed in MS Excelas follows.

D. Analysis of Data

The results of the assay with respect to the compounds exemplifiedherein are provided in the following table. A compound is consideredactive in RRA if it leads to at least a 1.5 fold increase in luciferaseactivity at 20 μM and is non-toxic, as determined by loss of signal(<0.75 fold increase). Fold increase is the amount of luciferaseactivity (measured in relative light units) over diluent control. SEMrepresents the standard error of the mean for fold increase (not shown).All compounds tested were found to be non-toxic. TABLE 23 Conc APPI FoldEx # (μg/mL) Increase Name  1 10 1.573-bromo-5-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]thiophene-2-sulfonamide  2 10 3.25-bromo-N-[(1S,2S)-1-(hydroxymethyl)-2- 3 2.34methylbutyl]thiophene-2-sulfonamide 10 3.06 10 4.26 1 1.79 20 4.5 20 5.520 6.3 3 2.96 1 1.54  3 10 14-bromo-5-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2- 20 1.5methylbutyl]thiophene-2-sulfonamide 20 25.4  4 10 3.35-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2- 3 2.98methylbutyl]thiophene-2-sulfonamide 3 3.52 20 4 20 4 20 5.4 20 3.3 205.1 1 2.22 10 5.03  5 3 1.1 2,5-dichloro-N-[(1S,2S)-1-(hydroxymethyl)-2-1 0.9 methylbutyl]thiophene-3-sulfonamide 0.3 0.9 10 1.1  6 10 1.14,5-dichloro-N-[(1S,2S)-1-(hydroxymethyl)-2- 20 1.5methylbutyl]thiophene-2-sulfonamide 20 2.9  7 20 3.2N-[(1S,2S)-1-(hydroxymethyl)-2- methylbutyl]thiophene-2-sulfonamide  810 3.7 5-chloro-N-[(1S)-1-(hydroxymethyl)-2- 3 2.5methylpropyl]thiophene-2-sulfonamide 20 3.3 20 5 20 3.8 1 1.5  9 10 1.815-bromo-N-[(1S)-1-(hydroxymethyl)-2- 20 4.1methylpropyl]thiophene-2-sulfonamide  10 20 1.25-bromo-N-[(1R)-1-(hydroxymethyl)-2- 20 6.4methylpropyl]thiophene-2-sulfonamide 20 1.7  11 20 24,5-dibromo-N-[(1S)-1-(hydroxymethyl)-2- 20 4.9methylpropyl]thiophene-2-sulfonamide  12 3 3.075-chloro-N-[(1S)-1-cyclohexyl-2- 1 2.58hydroxyethyl]thiophene-2-sulfonamide 10 4.2 20 4.3 3 3.1 20 8.6 1 2 103.9  13 10 3.94 5-bromo-N-[(1S)-1-cyclohexyl-2- 1 2.17hydroxyethyl]thiophene-2-sulfonamide 3 4.02  14 10 3.455-chloro-N-[1-(hydroxymethyl)-2- 3 1.87phenylpropyl]thiophene-2-sulfonamide 3 3.33  15 10 3.08 5-bromo-N-[ 1-(hydroxymethyl)-2- phenylpropyl]thiophene-2-sulfonamide  16 10 4.195-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2- 0.3 1.7methylbutyl]thiophene-2-sulfonamide 1 1.9 3 3.1 20 3.7 20 14.2 20 4.8 203.2 20 5.6 20 6.1 10 3.2 1 2.56 3 4.47  17 1 2.385-bromo-N-[(1S,2R)-1-(hydroxymethyl)-2- 3 3.47methylbutyl]thiophene-2-sulfonamide 10 3.29 20 6.1 20 3.5  18 10 5.235-bromo-N-[(1S,2S)-1-(hydroxymethyl)-2- 20 5.7methylbutyl]thiophene-2-sulfonamide 1,1-dioxide 3 2.19 20 3  19 20 65-chloro-N-[1-(hydroxymethyl)-2,3-dimethylpentyl]thiophene-2-sulfonamide  20 20 55-chloro-N-[1-(hydroxymethyl)-2- methylpentyl]thiophene-2-sulfonamide 21 20 3.3 5-chloro-N-[2-ethyl-1-(hydroxymethyl)hexyl]thiophene-2-sulfonamide  22 20 75-chloro-N-[2-hydroxy-1-(2,4,6-trimethylcyclohex-3-en-1-yl)ethyl]thiophene-2-sulfonamide  23 20 6.45-chloro-N-(1-cyclohex-3-en-1-yl-2- hydroxyethyl)thiophene-2-sulfonamide 24 20 6 5-chloro-N-(1-cyclopentyl-2-hydroxyethyl)thiophene-2-sulfonamide  25 20 7.5 5-bromo-N-[(1S)-1-(hydroxymethyl)-1,2-dimethylpropyl]thiophene-2-sulfonamide  26 20 55-chloro-N-[(1S)-1-(hydroxymethyl)-1,2-dimethylpropyl]thiophene-2-sulfonamide  27 20 8.75-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2,4-dimethylpentyl]thiophene-2-sulfonamide  28 20 75-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-(4-methoxyphenyl)propyl]thiophene-2-sulfonamide  29 20 2.65-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-methyloctyl]thiophene-2-sulfonamide  30 20 6.75-chloro-N-[(1S)-2-ethyl-1- 20 4.4(hydroxymethyl)butyl]thiophene-2-sulfonamide  31 20 4.45-chloro-N-[(1S,2R)-2-ethyl-1-(hydroxymethyl)-4-methylpentyl]thiophene-2-sulfonamide  32 20 55-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-(4-methoxyphenyl)butyl]thiophene-2-sulfonamide  33 20 6.55-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylpentyl]thiophene-2-sulfonamide  34 20 5.95-chloro-N-[(1S,2S)-2-ethyl-1-(hydroxymethyl)pentyl]thiophene-2-sulfonamide  35 20 3.75-chloro-N-[(1S,2R)-1-(hydroxymethyl)-4-methyl-2-propylpentyl]thiophene-2-sulfonamide  36 20 3.85-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-(4-methoxyphenyl)pentyl]thiophene-2-sulfonamide  37 20 4.85-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-propyloctyl]thiophene-2-sulfonamide  38 20 4.15-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-phenylpentyl]thiophene-2-sulfonamide  39 20 5.75-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylheptyl]thiophene-2-sulfonamide  40 20 3.95-chloro-N-[(1S)-2-propyl-1-(hydroxymethyl)pentyl]thiophene-2-sulfonamide  41 20 4.75-chloro-N-[(1S,2S)-2-ethyl-1-(hydroxymethyl)heptyl]thiophene-2-sulfonamide  42 20 3.45-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-isobutylheptyl]thiophene-2-sulfonamide  43 20 1.75-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-(4-methoxyphenyl)heptyl]thiophene-2-sulfonamide  44 20 1.65-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-pentyloctyl]thiophene-2-sulfonamide  45 20 2.15-chloro-N-[(1S,2R)-1-(hydroxymethyl)-2-phenylheptyl]thiophene-2-sulfonamide  46 20 5.85-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-phenylpropyl]thiophene-2-sulfonamide  47 20 5.45-chloro-N-[(1S,2S)-1-(hydroxymethyl)-4-methyl-2-phenylpentyl]thiophene-2-sulfonamide  48 20 4.75-chloro-N-[(1S,2R)-2-(2-furyl)-1-(hydroxymethyl)propyl]thiophene-2-sulfonamide  49 20 6.15-chloro-N-[(1S,2R)-2-(2-furyl)-1-(hydroxymethyl)butyl]thiophene-2-sulfonamide  50 20 7.75-chloro-N-[(1S,2R)-2-(2-furyl)-1-(hydroxymethyl)-4-methylpentyl]thiophene-2-sulfonamide  51 20 45-chloro-N-[(1S,2R)-2-(2-furyl)-1-(hydroxymethyl)octyl]thiophene-2-sulfonamide  52 20 75-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-isopropyloctyl]thiophene-2-sulfonamide  53 20 3.9N-[(1S,2S)-2-[1,1′-biphenyl]-4-yl-1-(hydroxymethyl)propyl]-5-chlorothiophene-2- sulfonamide  54 20 3.3N-[(1S,2S)-2-[1,1′-biphenyl]-4-yl-1-(hydroxymethyl)butyl]-5-chlorothiophene-2- sulfonamide  55 20 1.9N-[(1S,2S)-2-[1,1′-biphenyl]-4-yl-1-(hydroxymethyl)-4-methylpentyl]-5-chlorothiophene-2-sulfonamide  56 20 6.85-bromo-N-[(1S,2R)-1-(hydroxymethyl)-2,4-dimethylpentyl]thiophene-2-sulfonamide  57 20 6.45-bromo-N-[(1S,2R)-1-(hydroxymethyl)-2-methyloctyl]thiophene-2-sulfonamide  58 20 10 5-bromo-N-[(1S)-2-ethyl-1-(hydroxymethyl)butyl]thiophene-2-sulfonamide  59 20 4.55-bromo-N-[(1S,2R)-2-ethyl-1-(hydroxymethyl)-4-methylpentyl]thiophene-2-sulfonamide  60 20 6.95-bromo-N-[(1S,2S)-2-ethyl-1-(hydroxymethyl)pentyl]thiophene-2-sulfonamide  61 20 5.75-bromo-N-[(1S,2R)-1-(hydroxymethyl)-4-methyl-2-propylpentyl]thiophene-2-sulfonamide  62 20 4.55-bromo-N-[(1S,2S)-2-ethyl-1-(hydroxymethyl)heptyl]thiophene-2-sulfonamide  63 20 3.35-bromo-N-[(1S,2R)-1-(hydroxymethyl)-2-isobutylheptyl]thiophene-2-sulfonamide  64 20 2.95-bromo-N-[(1S,2R)-1-(hydroxymethyl)-2-(4-methoxyphenyl)heptyl]thiophene-2-sulfonamide  65 20 1.95-bromo-N-[(1S,2R)-1-(hydroxymethyl)-2-pentyloctyl]thiophene-2-sulfonamide  66 20 5.35-bromo-N-[(1S,2S)-1-(hydroxymethyl)-2-phenylpropyl]thiophene-2-sulfonamide  67 20 4.75-bromo-N-[(1S,2S)-1-(hydroxymethyl)-2-phenylbutyl]thiophene-2-sulfonamide  68 20 2.35-bromo-N-[(1S,2S)-1-(hydroxymethyl)-4-methyl-2-phenylpentyl]thiophene-2-sulfonamide  69 20 1.55-bromo-N-[(1S,2S)-1-(hydroxymethyl)-4-methyl-2-pyridin-3-ylpentyl]thiophene-2-sulfonamide  70 20 4.65-bromo-N-[(1S,2R)-2-(2-furyl)-1-(hydroxymethyl)propyl]thiophene-2-sulfonamide  71 20 5.25-bromo-N-[(1S,2R)-2-(2-furyl)-1-(hydroxymethyl)butyl]thiophene-2-sulfonamide  72 20 6.45-bromo-N-[(1S,2R)-2-(2-furyl)-1-(hydroxymethyl)-4-methylpentyl]thiophene-2-sulfonamide  73 20 1.95-bromo-N-[(1S,2R)-2-(2-furyl)-1-(hydroxymethyl)octyl]thiophene-2-sulfonamide  74 20 5.15-bromo-N-[(1S,2S)-2-ethyl-1-(hydroxymethyl)-3-methylbutyl]thiophene-2-sulfonamide  75 20 4.75-bromo-N-[(1S,2S)-1-(hydroxymethyl)-2-isopropyl-4-methylpentyl]thiophene-2-sulfonamide  76 20 3.2N-[(1S,2S)-2-[1,1′-biphenyl]-4-yl-1-(hydroxymethyl)butyl]-5-bromothiophene-2- sulfonamide  77A 20 3.15-chloro-N-[(1S,2R)-2-ethyl-1-(hydroxymethyl)octyl]thiophene-2-sulfonamide  77B 20 16.95-chloro-N-[(1S,2S)-2-ethyl-1-(hydroxymethyl)octyl]thiophene-2-sulfonamide  78A 20 5.85-bromo-N-[(1S,2R)-2-ethyl-1-(hydroxymethyl)octyl]thiophene-2-sulfonamide  78B 20 23.05-bromo-N-[(1S,2S)-2-ethyl-1-(hydroxymethyl)octyl]thiophene-2-sulfonamide  79 20 1.65-chloro-N-[(1S)-1-(hydroxymethyl)-2-(methylamino)butyl]-2-thiophenesulfonamide  80 20 1.85-chloro-N-[(1S)-2-(ethylamino)-2-(hydroxymethyl)propyl]-2-thiophenesulfonamide  81 20 1.55-chloro-N-[(1S)-2-[(2-hydroxyethyl)amino]-1-(hydroxymethyl)propyl]-2-thiophenesulfonamide  82 20 2.25-chloro-N-[(1S)-2-[(2-hydroxyethyl)amino]-1-(hydroxymethyl)butyl]-2-thiophenesulfonamide  83 20 1.75-chloro-N-[(1S)-2-[(2-hydroxyethyl)amino]-1-(hydroxymethyl)heptyl]-2-thiophenesulfonamide  84 20 3.8N-[(1S)-2-(benzylamino)-1-(hydroxymethyl)propyl]-5-chloro-2-thiophenesulfonamide  85 20 8.9N-[(1S)-2-(benzylamino)-1-(hydroxymethyl)butyl]-5-chloro-2-thiophenesulfonamide  86 20 205-chloro-N-[(1S)-2-(cyclopentylamino)-1-(hydroxymethyl)propyl]-2-thiophenesulfonamide  87 20 3.15-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-N-(2-phenoxyethyl)thiophene-2- sulfonamide  88 20 4.95-chloro-N-(3-chlorobenzyl)-N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]thiophene-2- sulfonamide  89 20 2.35-chloro-N-[(S)-2-hydroxy-1-phenylethyl]thiophene-2- sulfonamide  90 103.82 5-chloro-N-[(S)-1-(hydroxymethyl)-3- 20 3.6methylbutyl]thiophene-2-sulfonamide 3 1.74  91 20 25-chloro-N-[1-(hydroxymethyl)pentyl]thiophene-2- sulfonamide  92 20 1.55-chloro-N-(2-hydroxy-1,1-dimethylethyl)thiophene-2- sulfonamide  93 202 N-[1,1-bis(hydroxymethyl)propyl]-5-chlorothiophene- 2-sulfonamide  9410 2.4 5-chloro-N-[1-(hydroxymethyl)cyclopentyl]thiophene- 20 22-sulfonamide 20 3.4 20 2.2 20 2.2 20 5.8  95 10 1.95-chloro-N-[(S)-2-cyclohexyl-1- 20 4.1(hydroxymethyl)ethyl]thiophene-2-sulfonamide 20 3.8 20 1.9  96 20 1.5N-[(S)-1-benzyl-2-hydroxyethyl]-5-chlorothiophene-2- sulfonamide  97 201.8 5-chloro-N-[1-(hydroxymethyl)butyl]thiophene-2- sulfonamide  98 102.43 5-chloro-N-[(S)-1-(hydroxymethyl)-2,2- 20 2.8dimethylpropyl]thiophene-2-sulfonamide 20 4.9 20 2.4  99 20 1.55-chloro-N-[(R,R)-2-hydroxy-1-(hydroxymethyl)-2-(4-nitrophenyl)ethyl]thiophene-2-sulfonamide 100 20 1.55-chloro-N-[(S)-1-(hydroxymethyl)propyl]thiophene- 2-sulfonamide 101 202.7 N-[R-2-(benzylthio)-1-(hydroxymethyl)ethyl]-5-chlorothiophene-2-sulfonamide 102 20 1.6N-[(R,S)-2-(benzyloxy)-1-(hydroxymethyl)propyl]-5-chlorothiophene-2-sulfonamide 103 20 2.3 5-chloro-N-[(R,R)-2-hydroxy-1-(hydroxymethyl)propyl]thiophene-2-sulfonamide 104 20 2.55-bromo-N-[(S)-2-hydroxy-1-phenylethyl]thiophene-2- sulfonamide 105 103.93 5-bromo-N-[(S)-1-(hydroxymethyl)-3- 3 2.23methylbutyl]thiophene-2-sulfonamide 1 1.57 20 2.4 20 1.5 106 20 2.45-bromo-N-[1-(hydroxymethyl)pentyl]thiophene-2- sulfonamide 107 20 1.55-bromo-N-(2-hydroxy-1,1-dimethylethyl)thiophene-2- sulfonamide 108 20 2N-[1,1-bis(hydroxymethyl)propyl]-5-bromothiophene- 2-sulfonamide 109 102 5-bromo-N-[1-(hydroxymethyl)cyclopentyl]thiophene- 3 1.8 2-sulfonamide20 2.5 20 1.7 20 2.7 20 4.3 20 4.8 110 10 1.715-bromo-N-[(S)-2-cyclohexyl-1- 20 2.9(hydroxymethyl)ethyl]thiophene-2-sulfonamide 20 2.3 20 3.6 111 20 1.85-bromo-N-[(S)-1-(hydroxymethyl)-3-(methylthio)propyl]thiophene-2-sulfonamide 112 20 1.65-bromo-N-[1-(hydroxymethyl)butyl]thiophene-2- sulfonamide 113 10 2.865-bromo-N-[(S)-1-(hydroxymethyl)-2,2- 20 3.7dimethylpropyl]thiophene-2-sulfonamide 20 1.7 20 3 114 20 1.8N-[R-2-(benzylthio)-1-(hydroxymethyl)ethyl]-5-bromothiophene-2-sulfonamide 115 20 4.4 5-bromo-N-(R-2-hydroxy-1-{[(3-methylbenzyl)thio]methyl}ethyl)thiophene-2- sulfonamide 116 20 2.2N-{(S)-1-[4-(benzyloxy)benzyl]-2-hydroxyethyl}-5-bromothiophene-2-sulfonamide 117 20 2.6 5-bromo-N-[(R,R)-2-hydroxy-1-(hydroxymethyl)propyl]thiophene-2-sulfonamide 118 20 4.85-chloro-N-[(S,S)-1-formyl-2-methylbutyl]thiophene- 2-sulfonamide 119 104.1 5-chloro-N-[(S,S)-1-(1-hydroxyethyl)-2- 0.3 1.54methylbutyl]thiophene-2-sulfonamide 3 2.49 10 3.76 120 10 8.835-chloro-N-{(S,S)-1-[cyclopentyl(hydroxy)methyl]-2- 10 5.24methylbutyl}thiophene-2-sulfonamide 3 1.9 121 10 1.745-chloro-N-{(S)-2-hydroxy-1-[(S)-1- 10 1.5methylpropyl]octyl}thiophene-2-sulfonamide 10 1.56 122 10 1.685-chloro-N-{(S)-2-hydroxy-1-[(S)-1- 10 1.8methylpropyl]heptyl}thiophene-2-sulfonamide 123 10 2.145-chloro-N-{(S)-2-hydroxy-1-[(S)-1- 10 1.76methylpropyl]hexyl}thiophene-2-sulfonamide 124 10 2.325-chloro-N-{(S)-2-hydroxy-3-methyl-1-[(S)-1- 10 2.42methylpropyl]butyl}thiophene-2-sulfonamide 125 10 6.525-chloro-N-{(S)-2-hydroxy-3,3-dimethyl-1-[(S)-1- 3 1.58methylpropyl]butyl}thiophene-2-sulfonamide 10 3.94 126 10 5.255-chloro-N-{(S)-2-hydroxy-4-methyl-1-[(S)-1- 10 3.31methylpropyl]pentyl}thiophene-2-sulfonamide 127 10 4.275-chloro-N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]but-3-enyl}thiophene-2-sulfonamide 128 10 5.625-chloro-N-{(S)-2-hydroxy-1-[(S)-1- 1 1.52methylpropyl]pent-4-enyl}thiophene-2-sulfonamide 20 5.9 3 2.48 10 3.86129 10 7.43 5-chloro-N-{(S)-2-hydroxy-1-[(S)-1- 3 1.61methylpropyl]butyl}thiophene-2-sulfonamide 10 4.59 130 10 1.855-chloro-N-{(S,S)-1-[(4- fluorophenyl)(hydroxy)methyl]-2-methylbutyl}thiophene-2-sulfonamide 131 10 3.64 5-chloro-N-{(S,S)-1-[(4-chlorophenyl)(hydroxy)methyl]-2- methylbutyl}thiophene-2-sulfonamide 13210 1.68 5-chloro-N-{(S)-2-hydroxy-4-methyl-1-[(S)-1-methylpropyl]pent-3-enyl}thiophene-2-sulfonamide 133 10 2.515-chloro-N-{(S)-2-hydroxy-3-methyl-1-[(S)-1- 1.68methylpropyl]but-3-enyl}thiophene-2-sulfonamide 134 10 2.425-chloro-N-{(S,S)-1-[hydroxy(4-methoxyphenyl)methyl]-2-methylbutyl}thiophene-2- sulfonamide 135 10 1.935-chloro-N-{(S,E)-2-hydroxy-3-methyl-1-[(S)-1- 10 1.75methylpropyl]pent-3-enyl}thiophene-2-sulfonamide 136 10 2.135-chloro-N-{(S)-4-(1,3-dioxan-2-yl)-2-hydroxy-1-[(S)-1-methylpropyl]butyl}thiophene-2-sulfonamide 137 10 1.965-chloro-N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]hex-5-enyl}thiophene-2-sulfonamide 138 10 3.545-chloro-N-((S,S)-1-{hydroxy[4-(methylthio)phenyl]methyl}-2-methylbutyl)thiophene- 2-sulfonamide 139 103.97 5-chloro-N-{(S,S)-1-[[4- (dimethylamino)phenyl](hydroxy)methyl]-2-methylbutyl}thiophene-2-sulfonamide 140 10 2.57N-{(S,S)-1-[cyclopentyl(hydroxy)methyl]-2-methylbutyl}thiophene-2-sulfonamide 141 10 4.2N-{(S)-2-hydroxy-1-[(S)-1- methylpropyl]octyl}thiophene-2-sulfonamide142 10 3.59 N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]heptyl}thiophene-2-sulfonamide 143 10 1.64N-{(S)-2-hydroxy-1-[(S)-1- methylpropyl]hexyl}thiophene-2-sulfonamide144 10 1.51 N-{(S,S)-1-[hydroxy(2-methylphenyl)methyl]-2-methylbutyl}thiophene-2-sulfonamide 145 10 1.72N-{(S)-2-hydroxy-3,3-dimethyl-1-[(S)-1-methylpropyl]butyl}thiophene-2-sulfonamide 146 10 1.83N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]but-3-enyl}thiophene-2-sulfonamide 147 10 2.04N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]pent-4-enyl}thiophene-2-sulfonamide 148 10 1.52 N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]butyl}thiophene-2-sulfonamide 149 10 1.62N-{(S,S)-1-[hydroxy(4-methoxyphenyl)methyl]-2-methylbutyl}thiophene-2-sulfonamide 150 10 1.6N-{(S)-4-(1,3-dioxan-2-yl)-2-hydroxy-1-[(S)-1-methylpropyl]butyl}thiophene-2-sulfonamide 151 10 1.94N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]hex-5-enyl}thiophene-2-sulfonamide 152 10 1.51N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]pent-3-ynyl}thiophene-2-sulfonamide 153 10 2.09N-((S,S)-1-{hydroxy[4-(methylthio)phenyl]methyl}-2-methylbutyl)thiophene-2-sulfonamide 154 10 4.23 N-{(S,S)-1-[[4-dimethylamino)phenyl](hydroxy)methyl]-2-methylbutyl}thiophene-2-sulfonamide 155 20 4.25-chloro-N-{(S,S)-1-[(S)-cyclohex-2-en-1- 20 2.6yl(hydroxy)methyl]-2-methylbutyl}thiophene-2- sulfonamide 156 20 5.35-chloro-N-{(S,S,E)-2-hydroxy-1-[(S)-1- 20 5.9methylpropyl]hex-4-enyl}thiophene-2-sulfonamide 157 20 6.75-chloro-N-{(S,R,E)-2-hydroxy-1-[(S)-1- 20 6.8methylpropyl]hex-4-enyl}thiophene-2-sulfonamide 158 20 1.65-chloro-N-{(S,R,E)-2-hydroxy-1-[(S)-1- 20 5.4methylpropyl]hept-4-enyl}thiophene-2-sulfonamide 159 20 3.85-chloro-N-{(S,S)-2-hydroxy-4-methyl-1-[(S)-1- 20 3.2methylpropyl]pent-4-enyl}thiophene-2-sulfonamide 160 20 45-chloro-N-{(S,R)-2-hydroxy-4-methyl-1-[(S)-1- 20 3.3methylpropyl]pent-4-enyl}thiophene-2-sulfonamide 161 20 2.65-chloro-N-{(S,E)-2-hydroxy-1-[(S)-1-methylpropyl]-5-phenylpent-4-enyl}thiophene-2-sulfonamide 162 10 2.65-chloro-N-[(S,S)-1-(1-hydroxy-1-methylethyl)-2-methylbutyl]thiophene-2-sulfonamide 163 10 3.35-chloro-N-{(S)-2-hydroxy-1-[(S)-1-methylpropyl]-2-pentylheptyl}thiophene-2-sulfonamide 164 10 1.725-chloro-N-{(S,S)-1-[hydroxy(diphenyl)methyl]-2-methylbutyl}thiophene-2-sulfonamide 165 10 1.73N-{(S)-2-allyl-2-hydroxy-1-[(S)-1-methylpropyl]pent-4-enyl}-5-chlorothiophene-2-sulfonamide 166 10 1.785-chloro-N-{(S)-2-ethyl-2-hydroxy-1-[(S)-1-methylpropyl]butyl}thiophene-2-sulfonamide 167 10 3.42N-{(S,S)-1-[bis(4-chlorophenyl)(hydroxy)methyl]-2-methylbutyl}-5-chlorothiophene-2-sulfonamide 168 10 5.875-chloro-N-{(S)-2-hydroxy-2-isopropenyl-3-methyl-1- 10 1.6[(S)-1-methylpropyl]but-3-enyl}thiophene-2- 3 1.5 sulfonamide 169 101.51 5-chloro-N-((S,S)-1-{hydroxy[bis(4-methoxyphenyl)]methyl}-2-methylbutyl)thiophene-2- sulfonamide 170 101.95 5-chloro-N-{(S,E)-2-hydroxy-3-methyl-2-[(E)-1-methylprop-1-enyl]-1-[(S)-1-methylpropyl]pent-3-enyl}thiophene-2-sulfonamide 171 10 5.32N-{(S)-2-but-3-enyl-2-hydroxy-1-[(S)-1-methylpropyl]hex-5-enyl}-5-chlorothiophene-2- sulfonamide 172 10 25-chloro-N-((S,S)-1-{hydroxy[di(1-naphthyl)]methyl}-2-methylbutyl)thiophene-2-sulfonamide 173 10 1.55-bromo-N-{(S)-2-ethyl-2-hydroxy-1-[(S)-1-methylpropyl]butyl}thiophene-2-sulfonamide 174 10 1.755-bromo-N-{(S)-2-hydroxy-2-isopropenyl-3-methyl-1-[(S)-1-methylpropyl]but-3-enyl}thiophene-2- sulfonamide 175 10 2.275-bromo-N-{(S,E)-2-hydroxy-3-methyl-2-[(E)-1- 10 1.7methylprop-1-enyl]-1-[(S)-1-methylpropyl]pent-3- 20 3.3enyl}thiophene-2-sulfonamide 176 10 1.525-bromo-N-{(S)-2-but-3-enyl-2-hydroxy-1-[(S)-1-methylpropyl]hex-5-enyl}thiophene-2-sulfonamide 177 20 5.15-chloro-N-[1-(hydroxymethyl)cyclohexyl]thiophene- 20 3.7 2-sulfonamide178 20 2.3 5-chloro-N-[2-(hydroxymethyl)bicyclo[2.2.1]hept-2-yl]thiophene-2-sulfonamide 179 20 8.35-chloro-N-[1-(hydroxymethyl)-2,3-dihydro-H-inden-1-yl]thiophene-2-sulfonamide 180 20 2.35-chloro-N-[2-(hydroxymethyl)-2,3-dihydro-H-inden-2-yl]thiophene-2-sulfonamide 181 20 2.65-bromo-N-[1-(hydroxymethyl)cyclohexyl]thiophene- 2-sulfonamide 182 204.2 5-bromo-N-[2-(hydroxymethyl)bicyclo[2.2.1]hept-2-yl]thiophene-2-sulfonamide 183 20 3.75-bromo-N-[2-(hydroxymethyl)-2,3-dihydro-H-inden-2-yl]thiophene-2-sulfonamide 184 20 3.75-chloro-N-{(S,S)-1-[(S)-1-hydroxyethyl]-2- 20 2.7methylbutyl}thiophene-2-sulfonamide 185 20 2.45-chloro-N-{(S,S)-1-[R-1-hydroxyethyl]-2- 20 5.4methylbutyl}thiophene-2-sulfonamide 186 20 6.35-chloro-N-{(S,S)-2-hydroxy-1-[(S)-1- 20 2.4methylpropyl]pentyl}thiophene-2-sulfonamide 187 20 5.75-chloro-N-{(S,R)-2-hydroxy-1-[(S)-1- 20 3methylpropyl]pentyl}thiophene-2-sulfonamide 188 20 6.35-chloro-N-{(S,S)-2-hydroxy-1-[(S)-1- 20 3.6methylpropyl]pent-4-enyl}thiophene-2-sulfonamide 20 3.6 189 20 8.15-chloro-N-{(S,R)-2-hydroxy-1-[(S)-1- 20 7.9methylpropyl]pent-4-enyl}thiophene-2-sulfonamide 20 7.2 20 4.4 190 204.4 5-bromo-N-{(S,S)-1-[(S)-1-hydroxyethyl]-2- 20 2.6methylbutyl}thiophene-2-sulfonamide 191 20 5.45-bromo-N-{(S,S)-1-[R-1-hydroxyethyl]-2- 20 4.8methylbutyl}thiophene-2-sulfonamide 192 20 2.25-bromo-N-{(S,S)-2-hydroxy-1-[(S)-1- 20 1.9methylpropyl]pentyl}thiophene-2-sulfonamide 20 1.7 193 20 8.55-bromo-N-{(S,R)-2-hydroxy-1-[(S)-1- 20 4.5methylpropyl]pentyl}thiophene-2-sulfonamide 20 3 194 20 95-bromo-N-{(S,S)-2-hydroxy-1-[(S)-1- 20 5.3methylpropyl]pent-4-enyl}thiophene-2-sulfonamide 195 20 4.25-bromo-N-{(S,R)-2-hydroxy-1-[(S)-1- 20 6.7methylpropyl]pent-4-enyl}thiophene-2-sulfonamide 196 20 20.75-chloro-N-[(S,S)-2-methyl-1-(2,2,2-trifluoro-1-hydroxyethyl)butyl]thiophene-2-sulfonamide 197 20 3.45-chloro-N-[1-(1-hydroxybut-3- 20 1.8enyl)cyclohexyl]thiophene-2-sulfonamide 198 20 4.45-chloro-N-[1-(1-hydroxy-3-methylbut-3-enyl)cyclohexyl]thiophene-2-sulfonamide 199 20 2.85-chloro-N-[(S)-2-hydroxy-1-(4-methoxycyclohexyl)ethyl]thiophene-2-sulfonamide 199A  20 2.45-Chloro-N-[(S)-2-hydroxy-1-(4-hydroxycyclohexyl)ethyl]thiophene-2-sulfonamide 200 20 25-chloro-N-[(S)-2-hydroxy-1-(4-propoxycyclohexyl)ethyl]thiophene-2-sulfonamide 201 20 2.2N-{(S)-1-[4-(allyloxy)cyclohexyl]-2-hydroxyethyl}-5-chlorothiophene-2-sulfonamide 202 20 2N-{(S)-1-[4-(benzyloxy)cyclohexyl]-2-hydroxyethyl}-5-chlorothiophene-2-sulfonamide 203 20 1.5N-[1-acetyl-4-(hydroxymethyl)piperidin-4-yl]-5-chlorothiophene-2-sulfonamide 204 20 2.85-chloro-N-[(1S,2S)-1-(hydroxymethyl)-2- 20 2.5methylbutyl]-2-furansulfonamide 205 20 2.6N[(1S)-2-butyl-1-(hydroxymethyl)hexyl]-5-chloro-2- thiophenesulfonamide206 20 1.8 N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-furansulfonamide 207 20 5.6N-[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-5-iodo-2-thiophenesulfonamide 208 20 16.55-fluoro-N[(1S,2S)-1-(hydroxymethyl)-2-methylbutyl]-2-thiophenesulfonamide 209 20 2.14-[1-(5-chloro-thiophene-2-sulfonylamino)-2-hydroxy-ethyl]-piperidine-1-carboxylic acid tert-butyl ester 210 20 7.7N-[(1S,2S)-1-(hydroxymethyl)-2- methylbutyl]thiophene-2-sulfonamide 21120 1.9 5-chloro-N-[(S)-2-hydroxy-1-(4-benzylaminocyclohexyl)ethyl]thiophene-2- sulfonamide 212 20 1.75-chloro-N-[(S)-2-hydroxy-1-(4- methylaminocyclohexyl)ethyl]thiophene-2-sulfonamide 213 20 1.6 5-chloro-N-[(S)-2-hydroxy-1-(4-ethylaminocyclohexyl)ethyl]thiophene-2-sulfonamide 214 20 2.15-chloro-N-[(S)-2-hydroxy-1-(4-npropylaminocyclohexyl)ethyl]thiophene-2- sulfonamide 215 20 2.05-chloro-N-[(S)-2-hydroxy-1-(4-allylaminocyclohexyl)ethyl]thiophene-2-sulfonamide 216 20 1.55-chloro-N-[(S)-2-hydroxy-1-(4-(3-pyridyl)methylaminocyclohexyl)ethyl]thiophene-2- sulfonamide 217 20 1.85-chloro-N-[(S)-2-hydroxy-1-(4-morpholinocyclohexyl)ethyl]thiophene-2-sulfonamide 218 20 2.85-chloro-N-[(S)-2-hydroxy-1-(4-(4-pyridyl)methylaminocyclohexyl)ethyl]thiophene-2- sulfonamide 219 20 2.35-chloro-N-[(S)-2-hydroxy-1-(4-(2-pyridyl)methylaminocyclohexyl)ethyl]thiophene-2- sulfonamide 220 20 2.55-chloro-N-[(S)-2-hydroxy-1-(4-(carboethoxymethyl)aminocyclohexyl)ethyl]thiophene- 2-sulfonamide 221 204.9 5-chloro-N-[(S)-2-ethyl-1-formylbutyl]thiophene-2- sulfonamide 22220 7.2 5-chloro-N-[(S)-2-ethyl-1-(1- 9.7hydroxyethyl)butyl]thiophene-2-sulfonamide 223 20 5.65-chloro-N-[(S)-2-ethyl-1-(1-hydroxy-1- 8.4methylethyl)butyl]thiophene-2-sulfonamide 224 20 7.25-chloro-N-(2-hydroxy-1-tetrahydro-H-thiopyran-4-ylethyl)thiophene-2-sulfonamide 225 20 8.05-chloro-N-[(S)-2-hydroxy-1-piperidin-4- ylethyl]thiophene-2-sulfonamide226 20 7.6 N-[(S)-2-ethyl-1-(hydroxymethyl)butyl]thiophene-2-sulfonamide 227 20 26.3 N-[(S)-2-ethyl-1-(hydroxymethyl)butyl]-5-fluorothiophene-2-sulfonamide 228 20 9.95-chloro-N[(S)-1-(2,3-dihydro-H-inden-2-yl)-2-hydroxyethyl]thiophene-2-sulfonamide 229 20 6.35-chloro-N-{(S,S)-1-[(Z)-(hydroxyimino)methyl]-2-methylbutyl}thiophene-2-sulfonamide 230 20 4.85-chloro-N-{(S,S)-1-[(E)-(hydroxyimino)methyl]-2-methylbutyl}thiophene-2-sulfonamide

All publications cited in this specification are incorporated herein byreference. While the invention has been described with reference to aparticularly preferred embodiment, it will be appreciated thatmodifications can be made without departing from the spirit of theinvention. Such modifications are intended to fall within the scope ofthe appended claims.

1. A method of detecting an inhibitor of beta amyloid production,comprising assaying a sample from a subject treated with a compound offormula I by contacting said sample with an antibody to a compound offormula I, wherein said compound of formula I has the structure:

wherein: R₁ and R₂ are independently selected from the group consistingof hydrogen, alkyl, substituted alkyl, CF₃, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, phenyl, substituted phenyl, and (CH₂)_(n)(1,3)dioxane; n is2 to 5; R₃ is selected from the group consisting of hydrogen, alkyl, andsubstituted alkyl; R₄ is selected-from the group consisting of hydrogen,alkyl, substituted alkyl, alkylcycloalkyl, substituted alkylcycloalkyl,phenyl(substituted)alkyl, alkylOH, substituted alkylOH, alkylOBn,substituted alkylOBn, alkylpyridyl, substituted alkylpyridyl,alkylfuranyl, substituted alkylfuranyl, CH(OH)phenyl, CH(OH)substitutedphenyl, alkenyl, substituted alkenyl, cycloalkyl, substitutedcycloalkyl, N-substituted-piperidinyl, piperidinyl, substitutedpiperidinyl, tetrahydrothiopyran, substituted tetrahydrothiopyran,2-indane, substituted 2-indane, phenyl, substituted phenyl, alkylNHR₇,and substituted alkylNHR₇; with the proviso that R₃ and R₄ are not bothhydrogen; R₇ is alkyl, substituted alkyl, cycloalkyl, substitutedcycloalkyl, benzyl, substituted benzyl, alkylOH, substituted alkylOH,alkylSR₈, or substituted alkylSR₈; R₈ is alkyl, substituted alkyl,benzyl, or substituted benzyl; or R₃ and R₄ may be joined to form aring; R₅ is selected from the group consisting of hydrogen, lower alkyl,substituted lower alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, CH₂cycloalkyl, substituted CH₂cycloalkyl, benzyl,substituted benzyl, and CH₂CH₂QR₉; Q is O, NH or S; R₉ is lower alkyl,substituted lower alkyl, phenyl, or substituted phenyl; R₆ is selectedfrom the group consisting of hydrogen, halogen and CF₃; T is selectedfrom the group consisting of

W, Y and Z are independently selected from the group consisting of C,CR₁₀ and N; R₁₀ is selected from the group consisting of hydrogen andhalogen, with the proviso that at least one of W, Y and Z must be C; Xis selected from the group consisting of O, S, SO₂, and NR₁₁; R₁₁ isselected from the group consisting of hydrogen, lower alkyl, substitutedlower alkyl, benzyl, substituted benzyl, phenyl, and substituted phenyl;provided that when the compound contains one or more chiral centers, atleast one of the chiral centers must be of S-stereochemistry.
 2. Themethod according to claim 1, wherein said assay is a radioimmunoassay oran enzyme linked immunosorbent assay.
 3. A method of diagnosing acondition associated with abnormal levels of beta amyloid comprisingassaying a sample from a subject having abnormal levels of beta amyloidby contacting said sample with a compound of formula I, wherein saidcompound of formula I is of the structure:

wherein: R₁ and R₂ are independently selected from the group consistingof hydrogen, alkyl, substituted alkyl, CF₃, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, phenyl, substituted phenyl, and (CH₂)_(n)(1,3)dioxane; n is2 to 5; R₃ is selected from the group consisting of hydrogen, alkyl, andsubstituted alkyl; R₄ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkylcycloalkyl, substituted alkylcycloalkyl,phenyl(substituted)alkyl, alkylOH, substituted alkylOH, alkylOBn,substituted alkylOBn, alkylpyridyl, substituted alkylpyridyl,alkylfuranyl, substituted alkylfuranyl, CH(OH)phenyl, CH(OH)substitutedphenyl, alkenyl, substituted alkenyl, cycloalkyl, substitutedcycloalkyl, N-substituted-piperidinyl, piperidinyl, substitutedpiperidinyl, tetrahydrothiopyran, substituted tetrahydrothiopyran,2-indane, substituted 2-indane, phenyl, substituted phenyl, alkylNHR₇,and substituted alkylNHR₇; with the proviso that R₃ and R₄ are not bothhydrogen; R₇ is alkyl, substituted alkyl, cycloalkyl, substitutedcycloalkyl, benzyl, substituted benzyl, alkylOH, substituted alkylOH,alkylSR₈, or substituted alkylSR₈; R₈ is alkyl, substituted alkyl,benzyl, or substituted benzyl; or R₃ and R₄ may be joined to form aring; R₅ is selected from the group consisting of hydrogen, lower alkyl,substituted lower alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, CH₂cycloalkyl, substituted CH₂cycloalkyl, benzyl,substituted benzyl, and CH₂CH₂QR₉; Q is O, NH or S; R₉ is lower alkyl,substituted lower alkyl, phenyl, or substituted phenyl; R₆ is selectedfrom the group consisting of hydrogen, halogen and CF₃; T is selectedfrom the group consisting of

W, Y and Z are independently selected from the group consisting of C,CR₁₀ and N; R₁₀ is selected from the group consisting of hydrogen andhalogen, with the proviso that at least one of W, Y and Z must be C; Xis selected from the group consisting of O, S, SO₂, and NR₁₁; R₁₁ isselected from the group consisting of hydrogen, lower alkyl, substitutedlower alkyl, benzyl, substituted benzyl, phenyl, and substituted phenyl;provided that when the compound contains one or more chiral centers, atleast one of the chiral centers must be of S-stereochemistry.
 4. Themethod according to claim 3, wherein said condition is selected from thegroup consisting of Alzheimer's Disease, amyloid angiopathy, cerebralamyloid angiopathy, systemic amyloidosis, hereditary cerebral hemorrhagewith amyloidosis of the Dutch type, inclusion body myositis, and Down'ssyndrome.
 5. The method according to claim 3, wherein said assay is anenzyme linked immunosorbent assay.
 6. The method according to claim 3,wherein said assay is a radioimmunoassay.
 7. The method according toclaim 3, wherein said subject had been administered a compound offormula I.
 8. The method according to claim 3, wherein said assayfurther comprises contacting said sample with an antibody to a compoundof formula I.
 9. A method of preparing a compound of the structure:

wherein: R₁ and R₂ are independently selected from the group consistingof hydrogen, alkyl substituted alkyl, CF₃, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,phenyl, substituted phenyl, and (CH2)_(n)(1,3)dioxane; n is 2 to 5; R₃is selected from the group consisting of hydrogen, alkyl, andsubstituted alkyl; R₄ is selected from the group consisting of hydrogen,alky, substituted alkyl, alkylcycloalkyl, substituted alkylcycloalkyl,phenyl(substituted)alkyl, alkylOH, substituted alkylOH, alkylOBn,substituted alkylOBn, alkylpyridyl, substituted alkylpyridyl,alkylfuranyl, substituted alkylfuranyl, CH(OH)phenyl, CH(OH)substitutedphenyl, alkenyl, substituted alkenyl, cycloalkyl, substitutedcycloalkyl, N-substituted-piperidinyl, piperidinyl, substitutedpiperidinyl, tetrahydrothiopyran, substituted tetrahydrothiopyran,2-indane, substituted 2-indane, phenyl, substituted phenyl, alkylNHR₇,and substituted alkylNHR₇; with the proviso that R₃ and R₄ are not bothhydrogen; R₇ is alkyl, substituted alkyl, cycloalkyl, substitutedcycloalkyl, benzyl, substituted benzyl, alkylOH, substituted alkylOH,alkylSR₈, or substituted alkylSR₈; R₈ is alkyl, substituted alkyl,benzyl, or substituted benzyl; or R₃ and R₄ may be joined to form aring; R₅ is selected from the group consisting of hydrogen, lower alkyl,substituted lower alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, CH₂cycloalkyl, substituted CH₂cycloalkyl, benzyl,substituted benzyl, and CH₂CH₂QR₉; Q is O, NH or S; R₉ is lower alkyl,substituted lower alkyl, phenyl, or substituted phenyl, R₆ is selectedfrom the group consisting of hydrogen, halogen and CF₃; W, Y and Z areindependently selected from the group consisting of C, CR₁₀ and N; R₁₀is selected from the group consisting of hydrogen and halogen, with theproviso that at least one of W, Y and Z must be C; X is selected fromthe group consisting of O, S, SO₂, and NR₁₁; R₁₁ is selected from thegroup consisting of hydrogen, lower alkyl, substituted lower alkyl,benzyl, substituted benzyl, phenyl, and substituted phenyl; providedthat when the compound contains one or more chiral centers, at least oneof the chiral centers must be of S-stereochemistry; said methodcomprising reacting a 2-aminoalcohol of the structure:

with a sulfonyl halide of the structure:


10. The method according to claim 9, wherein when R₁ and R₂ are H, themethod further comprises oxidizing said compound to an aldehyde of thestructure:


11. The method according to claim 10, further comprising alkylating saidaldehyde to a secondary alcohol.
 12. The method according to claim 11,wherein the secondary alcohol is of the structure: